Note: Descriptions are shown in the official language in which they were submitted.
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WO 98104~37 PCTIUS97/13422-
NOVEL COMPOUNDS AND COMPOSITIONS FOR TREAT~NG DISEASES ASSOCIATED
WITH TRYPTASE ACTIVITY
~ 3 This application claims the benefit of U.S. Provisional Application No. 60/023,139, filed
July 30, 1996.
Field of the Invention:
6 This invention relates to novel methods and compositions for treating diseases associated
with tryptase activity by ~lmini~tration of novel tryptase inhibitors.
Description of the Field:
g Tryptase, the predominant protease secreted from human mast cells, is thought to be
involved in neuropeptide processing and tissue infl~mm~tion Tryptase concentrations are
elevated in the bloodstream for several hours following anaphylaxis (Schwartz el al. (1987)
12 N. ~ng. J. Med. 316:1622-1626), are increased in nasal and lung lavage fluid from atopic
subiects following specific antigen challenge (Castells e~ al. (198~) J. Allerg. Clin. ~mmunol.
141 :563-568) and are elevated in lung lavage fluid of atopic asthmatics after endobronchial
allergen challenge. Smokers often have striking elevations of bronchoalveolar lavage fluid
tryptase levels, a finding that provides some support for the hypothesis that release of proteinase
from activated mast cells could contribute to lung destruction in smoker's emphysema.
18 (Celenteron et ~1. (198~) Chest 94: 119-123). In addition, tryptase has been shown to be a potent
mitogen for fibroblasts, suggesting that it is involved in pulmonary fibrosis and interstitial lung
disease (Ross e~ al. (l991)J. Clln. Invest. 88:493-499).
21 Asthma is recognized as an infl~mm~tory disorder (Hood et al. (1984)
In: Benjarnin-CIlmming.~, ed. Immunology 2nd ed.) and frequently is characterized by
progressive development of hyper responsiveness of the trachea and bronchi to both
24 imrnunospecific allergens and generalized chemical or physical stimuli. The disease involves
~ multiple biochemical mediators in both its acute and chronic stages. The hyper responsiveness of
asthmatic bronchiolar tissue is believed to be the result of chronic infl~mm~tory reactions, which
27 irritate and damage the epithelium lining the airway wall and promote pathological thickening of
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W O 98t04537 PCTrUS97113422-
the underlying tissue. Bronchial biopsies in patients with only mild asthma have features of
infl~mm~tion in the airway wall.
3 Allergic responses to inhaled allergens can initiate the infl~n~m~tory sequence. For
example, allergens can activate mast cells and basophils~ which are present in the epithelium and
underlying smooth muscle tissue by binding IgE located on the cell surface. Activated mast cells
6 release a number of preformed or primary chemical mediators (e.g., histamine) of the
infl~3mm~tory response and generate numerous other secondary mediators of infl~mm~tion
(e.g., superoxide, lipid derived mediators, etc.) in situ. In addition, several large molecules (e.g.,
g proteoglycans, tryptase, chymase, etc.) are released by degranulation of mast cells.
The release of these preforrned mediators from mast cells probably accounts for the early
bronchiolar constriction in the asthmatic reaction to air borne allergens. The early phase of the
2 asthmatic reaction peaks approximately fifteen minutes after exposure to allergen and is
generally followed by recovery over the ensuing one to two hours. Twenty five to thirty five
percent of the patient population experience a further decline in respiratory function which
m~ximi7es six to twelve hours after exposure. This late reaction phase is accompanied by a
marked increase in the number of infl~mm~tl~ry cells (e.g., eosinophils, neutrophils,
lymphocytes, etc.) infiltrating the bronchiolar tissue. The infiltrating cells are attracted to the site
18 by release of mast cell derived chemotactic agents and then become activated during the late
reaction phase. The late asthmatic response is believed to be a secondary infl~mm~tory reaction
merli~t~d in part by the secretory activity o~ granulocytes.
21 Tryptase is implicated in the degradation of vasodilating and bronchorelaxing
neuropeptides (Caughey et al. (1988) ~Pharmacol. ~xp. Ther. 244:133-137; Franconi et al.
(1988)J: Pharmacol. Exp. Ther. 248:947-951;andTam etal. (1990) Am. J. Respir. CellMol.
24 Biol. 3:27-32) and modulation of bronchial responsiveness to histamine (Sekizawa et al. (1989)
Clin. Invest. 83: 175- 179). These findings suggest that tryptase may increase
bronchoconstriction in asthma by destroying bronchodilating peptides. Tryptase cleaves
27 fibrinogen o~-chains and high molecular weight kinninogen, which suggests that tryptase plays a
role with heparin as a local anticoagulant. Tryptase activates prostromelysin (pro-MMP-3) and
procollagenase ~pro-MMP-1) via MMP-3, which suggests that tryptase is involved in tissue
infl~n~m~tion and remodeling and joint destruction in rheumatoid arthritis. Further,
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~mini.ctration of tryptase inhibitor protects against development of the late and airway hyper
responsive phases in allergen challenged sheep (Clark et al. (1995) A~. J: ~espir. Cr~t. Care
3 Med. 152: 2076-2083) and inhibits the immediate cutaneous response to intraderrnal injection of
- allergen in allergic sheep (Molinari et al. (1995) ,4mer. Physiol. Soc. 79(6):1966-1970). All of
the above-described fimdings clearly indicate the applicability of tryptase inhibitors as therapeutic
6 agents in treating asthma and other disorders associated with infl~mm~tion of the respiratory
tract.
The disclosures of these and other documents referred to throughout this application are
g incorporated herein by reference.
SUMMARY OF THE INVENTION
This application relates to a compound of Formula I:
1 2Rl-xl-x2-x3-x4\
R2 X9 X8 X7 X6 /
in which:
X5is (C3 ,4)cycloalkylene, hetero(C3.,4)cycloalkylene, (C6 ,4)arylene or
hetero(Cs,4)arylene;
X4 and x6 are independently (C0 2)alkylene;
X' and X9 are independently a covalent bond, -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R3)-,
1 8 -N(R3)C(o)-, -S(O)2N(R3)-, -N(R3)S(o)2-, -OC(O)N(R3)-, -N(R3)C(O)O-, -N(R3)C(o)N(R3)- or
~ -OC(O)O-, wherein each R3 is independently hydrogen, (Cl 3)alkyl or (C3 8)cycloalkyl, with the
proviso that Xl and X9 are not both covalent bonds;
21X3 and X7 are independently -C(O)-, -C(O)O-, -OC(O)-, -C(O)N(R3)-, -N(R3)C(o)-,
-S(O)2N(R3)-, -N(R3)S(o)2-, -OC(O)N(R3)-, -N(R3)C(O)O-, -N(R3)C(O)N(R3)- or -OC(O)O-,
wherein R3 is as defined above;
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X2 and x8 are independently (C, 8)alkylene, hetero(C, 8)alkylene, -X'~-X"- or -X'l-X10-,
wherein X~~ is (CO.4)alkylene or hetero(C3.4)alkylene and X~ ~ is (C3.8)cycloalkylene or
3 hetero(C3 8)cycloalkylene;
R' is R4-X'2- or R5-X'3-, wherein:
R4 is arnino, arnidino, guanidino, I-iminoethyl or methylarnino,
6 X'2 is ~C4 6)alkylene, hetero(C4.6)alkylene, heterooxo(C4 6)alkylene,
oxo(C4 6)alkylene or -X'4-X'5-X'6-, wherein X'5is(C36)cycloalkylene,
hetero(Cs 6)arylene, hetero(C3 6)cycloalkylene or phenylene, X~4is (Cn,4)alkylene and x~6
g is (Cnl6)alkylene, wherein the surn of nl4 and nl6 is 0, 1, 2, 3 or 4,
R5 is a group selected from azetidin-3-yl, benzoimidazol-4-yl, benzoimidazol-S-yl
imidazol-l-yl, Tmicl~ol-2-yl, imidazol-4-yl, 2-imidazolin-2-yl, 2-imidazolin-3-yl,
12 2-methylimidazol- 1 -yl, 4-methylimidazol- 1 -yl, 5-methylimidazol- 1 -yl,
l-methylpiperid-3-yl, 1-methylpiperid-4-yl, piperid-3-yl, piperid-4-yl, piperazin-l-yl,
piperazin-2-yl, pyrid-3-yl, pyrid-4-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrrolidin-3-yl,
1,4,5,6-tetrahyd~o~y~ idin-2-yl, 1,4,5,6-tetrahydropyrimidin-4-yl and
1,4,5,6-tetrahydlv~y~ idin-5-yl and any carbocyclic ketone or thioketone derivative
thereof, which group is optionally substituted with one or more radicals selected from
18 halo, hydroxy, mercapto, (C, 8)alkyl, (C3 ,4)cycloalkyl, (C6 ,4)aryl, (C6 ,4)aryl(C, 4)alkyl,
(C, 8)alkanoyl, (C, 8)alkyloxy, (C6 ,4)aryloxy, (C3 ,4)cycloalkyloxy, (Cl 4)alkyloxy,
(C, 8)alkylthio, (C3 ,4)cycloalkylthio, (C6 ,4)arylthio and -NR6R7, wherein R6 and R7 are
21 independently selected from hydrogen, (C, 8)alkyl, (C, 8)alkanoyl, (C3 ,4)cycloalkyl or
(C6 ,4)aryl and
X~3 is (CO 6)alkylene, hetero(C2 6)alkylene, heterooxo(C3 6)alkylene,
24 oxo(C2 6)alkylene or -X~7-X~8-X~9-, wherein Xl8 is as defined above for X'5, X~7is
(Cn~7)alkylene and x~8 is (Cn,8)alkylene, wherein the sum of nl 7 and nl 8 is 0, 1 or 2; and
R2 is R8-X20- or RY-X2'-, wherein:
27 R8 is arnino, l-iminoethyl or methylarnino,
X20 is (C4 6)alkylene, hetero(C4.6)alkylene, heterooxo(C4 6)alkylene,
oxo(C4 6)alkylene or -X22-X23-X24-, wherein X23 is as defined above for X~5,X22is
(Cn22)alkylene and X24 is (Cn24)alkylene, wherein the sum of n22 and n24 is 0, 1, 2, 3 or 4,
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WO 98/04537 PCT/US97/13422
with the proviso that when R~ is amino then X22 is not (C4 6)alkylene or oxa(C4 ~)alkylene
and n22 is not l, 2, 3 or 4,
3 R9 is as defined above for Rs and
X2' is (CO.6)alkylene, hetero(C2 6)alkylene, heterooxo(C3 6)alkylene,
oxo(C2 6)alkylene or -X25-X26-X2'-, wherein X26 is as defined above for X'5,X25is
6 (Cn25)alkylene and X27 is (Cn2~)alkylene, wherein the sum of n25 and n27 is 0, l or 2;
wherein each alkylene, cycloalkylene, heteroalkylene, heterocycloalkylene, phenylene,
arylene and heteroarylene, as defined above, are optionally substituted with one or more
g radicals selected from halo, hydroxy, mercapto, (Cl 8)alkyl, (C3 l4)cycloalkyl, (C6 l4)aryl,
(C6 l4)aryl(C, 4)alkyl, (Cl 8)alkanoyl, (C, 8)alkyloxy, (C6 l4)aryloxy, (C3 ,4)cycloalkyloxy,
(Cl 4)alkyloxy, (Cl 8)alkylthio, (C3 l4)cycloalkylthio, (C6 l4)arylthio and -NR6R7, wherein
12 R6 and R7 are as defined above; with the proviso that covalent bonds do not occur
between heteroatoms contained within R', X2,X4,X6,X8 and R~ and any heteroatoms
contained with X3,X5, X7 and X9; and
the phannaceutically acceptable salts, N-oxides, prodrug derivatives and protected derivatives
thereof.
A second aspect of this application relates to a compound of Formula I:
8 R]-xl-x2-x3-x4\
R2 X9 X8 X7 x6~
in which:
X4-X5-X6 together are (C2 l2)alkylene or hetero(C3 12)alkylene;
21 X~ and X9 are independently a covalent bond, -C(O)-, -C(O)O-, -OC(O~-, -C(o)N(R3)-,
-N(R3)C(o)-, -S(o)2N(R3)-, -N(R3)S(o)2-, -OC(O)N(R3)-, -N(R3)C(o)o-, -N(R3)C(O)N(R3)- or
-OC(O)O-, wherein each R3 is independently hydrogen, (Cl 3)alkyl or (C3 8)cycloalkyl, with the
24 proviso that X' and X9 are not both covalent bonds;
X3 and X7 are independently -C(O)-, -C~O)O-, -OC(O)-, -C(O)N(R3)-, -N(R3)C(o)-~
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-S(0)2N(R3)-, -N(R3)S(o)2-, -OC(o)N(R3)-, -N(R3)C(O)O-, -N(R3)C(O)N(R3)- or -OC(O)O-,
wherein R3 is as defined above;
3 X2 and x8 are independently (C, 8)alkylene, hetero(C, 8)alkylene, -X'~-X"- or -X"-X'~-,
wherein X~~ is (C0 4)alkylene or hetero~C3.4)alkylene and X~ I is (C3.~)cycloalkylene or
hetero(C3 8)cycloalkylene,
6 R~ is R4-XI2- or R5-XI3-, wherein:
R4 is arnino, amidino, guanidino, 1-iminoethyl or methylamino,
Xl2 is (C4.6)alkylene, hetero(C4.6)alkylene, heterooxo(C4 6)alkylene,
g oxo(C4 6)alkylene or -Xl4-X~5-X~6-, wherein Xl5 is (C3 6)cycloalkylene,
hetero(C5 6)arylene, hetero(C3 6)cycloalkylene or phenylene, X14 iS (Cnl4)alkylene and Xl6
is (Cnl6)alkylene, wherein the sum of nl4 and nl 6 is 0, 1, 2, 3 or 4,
12 R5 is a group selected from azetidin-3-yl, benzoimidazol-4-yl, benzoimidazol-5-yl
imidazol-l-yl, imidazol-2-yl, imidazol-4-yl, 2-imidazolin-2-yl, 2-imidazolin-3-yl,
2-methylimidazol- 1 -yl, 4-methylimidazol- 1 -yl, 5-methylimidazol- l -yl,
1-methylpiperid-3-yl, l-methylpiperid-4-yl, piperid-3-yl,piperid-4-yl,piperazin-1-yl,
piperazin-2-yl, pyrid-3-yl, pyrid-4-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyrrolidin-3-yl,
1,4,5,6-tetrahydropyrimidin-2-yl, 1,4,5,6-tetrahydropyrimidin-4-yl and
8 1,4,5,6-tetrahydropyrimidin-5-yl and any carbocyclic ketone or thioketone derivative
thereof, which group is optionally substituted with one or more radicals selected from
halo, hydroxy, mercapto, (C, 8)alkyl, (C3.,4)cycloalkyl, (C6.,4)aryl, (C6.l4)aryl(CI 4)alkyl,
21 (Cl.8)alkanoyl, (Cl.8)alkyloxy, (C6 ~4)aryloxy, (C3 l4)cycloalkyloxy, (C, 4)alkyloxy,
(C~ 8)alkylthio, (C3 ~4)cycloalkylthio, (C6 l4)arylthio and -NR6R7, wherein R6 and R7 are
independently selected from hydrogen, (Cl 8)alkyl, (Cl.8)alkanoyl, (C3 l4)cycloalkyl or
24 (C6 ]4)aryl and
X'3 is (C0 6)alkylene, hetero(C2 6)alkylene, heterooxo(C3 6)alkylene,
oxo(C2 6)alkylene or -Xl7-X'8-X~9-, wherein Xl8 is as defined above for X15, X17 iS
27 (Cn~)alkylene and Xl8 iS (Cn,8)alkylene, wherein the sum of n l 7 and nl 8 is 0, 1 or 2; and
R7 is R8-X20- or R9-X2l-, wherein:
R8 is as defined above for R4,
X20 is (C4 6)alkylene, hetero(C4 6)alkylene, heterooxo(C4 6)alkylene,
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oxo(C4 6)alkylene or -X22-X23-X24-, wherein X23 is as defined above for X'5, X22 iS
(Cn22)alkylene and X24 is (Cn24)alkylene, wherein the sum of n22 and n24 is 0, 1, 2, 3 or 4,
3 R9 is as defined above for Rs and
X2' is (CO.6)alkylene, hetero(C2 6)alkylene, heterooxo(C3 6)alkylene,
oxo(C2 6)al~ylene or -X25-X26-X27-, wherein X26 is as defined above for X15, X25 iS
6 (Cn25)alkylene and X27 is (Cn27)alkylene, wherein the sum of n25 and n27 is 0, l or 2;
wherein each alkylene, cycloalkylene, heteroalkylene, heterocycloalkylene, phenylene,
arylene and heteroarylene, as defined above, are optionally substituted with one or more
g radicals selected from halo, hydroxy, mercapto, (C~ 8)alkyl, (C3 ~4)cycloalkyl, (C6.~4)aryl,
(C6 ,4)aryl(C, 4)alkyl, (C, 8)alkanoyl, (C, 8)alkyloxy, (C6 l4)aryloxy, (C3 l4)cycloalkyloxy,
(C, 4)alkyloxy, (C, ~)alkylthio, (C3 ,4)cycloalkylthio, (C6 ,4)arylthio and -NR~R7, wherein
12 R6 and R7 are as defined above; with the proviso that covalent bonds do not occur
between heteroatoms contained within R~, X2, X4, X6, X8 and R2 and any heteroatoms
contained with X~, X~, X7 and X9; and
15 the pharmaceutically acceptable salts, N-oxides, prodrug derivatives and protected derivatives
thereof.
A third aspect of this invention is a pharmaceutical composition which contains a
18 compound of Formula I, or a ph~ c.eutically acceptable salt~ oxide or prodrug derivative
thereof in admixture with one or more suitable excipients.
A fourth aspect of this invention is a method of treating a disease in an animal in which
21 tryptase activity contributes to the pathology and/or symptomatology of the disease, which
method comprises ~lmini~tering to the animal a therapeutically effective amount of compound of
Formula I or a ph~ reutically acceptable salt, N-oxide or prodrug derivative thereof.
24 A fifth aspect of this invention is the processes for preparing compounds of Formula I
and the ph~ ceutically acceptable salts, ~-oxides, prodrug derivatives and protected
derivatives thereof as set forth in "Detailed Description of the Invention".
27 DETAILED DESCRIPTION OF THE INVENTION
Definitions:
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Unless otherwise stated, the following terms used in the specification and claims have the
meanings given below:
3 "Alkanoyl" means the radical -C(O)R, wherein R is alkyl as defined below, having
overall the number of carbon atoms indicated (e.g., (C, g)alkanoyl includes the radicals formyl,
acetyl, propionyl, butyryl, isobutyryl, crotonoyl, isocrotonyl, etc.).
6 "Alkyl", as in alkyl, arylalkyl, alkyloxy, alkylthio, means a straight or branched, saturated
or unsaturated hydrocarbon radical having the number of carbon atoms indicated (e.g.,
(Cl 8)alkyl includes methyl, ethyl, propyl, isopropyl, hutyl, sec-butyl, isobutyl, ~ert-butyl, vinyl,
g allyl, l-propenyl, isopropenyl, l-butenyl, 2-butenyl, 3-butenyl, 2-methylallyl, ethynyl,
1-pl~yllyl, 2-propynyl, etc.).
"Alkylene" means a straight, saturated or unsaturated hydrocarbon divalent radical having
12 the number of carbon atoms indicated (e.g., (CO 6)alkylene includes methylene (-CH2-), ethylene
(-(CH2)2-), vinylene (-CH:CH-), ethynylene (-C C-), 2-propylene (-CH:CH CH2-), l-propylene
(-CH2-CH:CH-), tetramethylene (-(CH2)4-), pentamethylene (-(CH,)s-) and hexamethylene
15 (-(CH2)6-), etc.). The term (C0)alkylene is meant to represent a covalent bond.
"Alkyloxy" means the radical -OR, wherein R is alkyl as defined above, having the
number of carbon atoms indicated (e.g., (C, 8)alkyloxy includes the radicals methoxy, ethoxy,
18 propoxy, isopropoxy, butoxy, isobutoxy, etc.).
"Alkylthio" means the radical -SR, wherein R is alkyl as defined above, having the
number of carbon atoms indicated (e.g., (Cl 8)alkylthio includes the radicals methylthio,
21 ethyl~io, propylthio, isopropylthio, butylthio, isobutylthio, etc.~.
"Animal" includes humans, non-human m~mm~l~ (e.g., dogs, cats, rabbits, cattle, horses,
sheep, goats, swine, deer, etc.) and non-m~mm~l~ (e.g., birds, etc.).
24 "Aryl", as in aryl, arylalkyl, aryloxy and arylthio, means an aromatic monocyclic or
polycyclic hydrocarbon radical cont~ining the number of carbon atoms indicated, wherein the
carbon atom with the free valence is a member of an aromatic ring, and any carbocylic ketone or
27 thioketone derivative thereof (e.g., (C6 l4)aryl includes phenyl, naphthyl, anthracenyl,
phe~ hrcnyl, 1,2,3,4-tetrahydronaphth-5-yl, 1-oxo-1,2-dihydronaphth-6-yl,
I-thioxo-1,2-dihydronaphth-7-yl, etc.).
"Arylene" means an aromatic monocyclic or polycyclic hydrocarbon divalent radical
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W O 98104S37 PCT~US97/13422
conS~inin~ the number of carbon atoms indicated, wherein the carbon atoms with the free valence
are members of an aromatic ring, and any carbocylic ketone or thioketone derivative thereof
3 (e.g., ~C6 ,4)arylene includes 1,4-phenylene, 1,3-phenylene, 1,4-naphthylene, 2,6-naphthylene,
I,4-anthracenylene, 2,6-anthracenylene, 1,6-phenanthrenylene,
1,2,3,4-tetrahydro-5,8-naphthylene, 1-oxo-1,2-dihydro-5,7-naphthylene,
6 1 -thioxo- 1 ,2-dihydro-5,8-naphthylene, etc.).
"Aryloxy" means the radical -OR, wherein R is aryl, as defined above, having the number
of carbon atoms indicated (e.g., (C6 ,4)aryloxy includes the radicals phenoxy, naphthyloxy,
9 anthracenyloxy, etc.).
"Arylthio" means the radical -SR, wherein R is aryl, as defined above, having the number
of carbon atoms indicated (e.g., (C6 ,4)arylthio includes the radicals phenylthio, naphthylthio,
12 anthracenylthio, etc.).
"Cycloalkyl", as in cycloalkyl and cycloalkyloxy, means a saturated or unsaturated,
monocyclic or polycyclic hydrocarbon radical containing the number of carbon atoms indicated,
5 wherein the carbon atom with the free valence is a member of a non-aromatic ring, and any
carbocyclic ketone and thioketone derivative thereof (e.g., (C3 l4)cycloalkyl includes
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2,5-cyclohexadienyl,
18 bicyclo[2.2.2]octyl, 1,2,3,4-tetrahydronaphth-1-yl, oxocyclohexyl, dioxocyclohexyl,
thiocyclohexyl, etc.).
"Cycloalkylene" means a saturated or unsaturated, monocyclic or polycyclic hydrocarbon
2 1 divalent radical co~ ng the number of carbon atoms indicated, wherein the carbon atoms with
the free valence are members of a non-aromatic ring, and any carbocyclic ketone and thioketone
derivative thereof (e.g., (C3 6)cycloalkylene includes 1,2-cyclopropylene, 1 ,2-cyclobutylene,
24 1,3-cyclobutylene, 1,2-cyclopentylene, 1,3-cyclopentylene, 1,4-cyclopentylene,
1,4-cyclohexylene, 3-cyclohexen-1,2-ylene, 2,5-cyclohexadien-1,4-ylene,
1 ,4-bicyclo[2.2.2]octylene, 1,2 ,3 ,4-tetrahydro- 1 ,4-naphthylene, 5-oxo- 1 ,3-cyclohexylene,
27 2,5-dioxo-1,4-cyclohexylene, 5-thioxo-1,4-cyclohexylene, etc.).
"Cycloalkyloxy" means the radical -OR, wherein R is cycloalkyl, as defined above,
having the number of carbon atoms indicated (e.g., (C3 ,4)cycloalkyloxy includes the radicals
30 cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, etc.).
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-10-
"Cycloalkylthio" means the radical -OR, wherein R is cycloalkyl, as defined above,
having the number of carbon atoms indicated (e.g., (C3 ,4)cycloalkylthio includes the radicals
3 cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, etc.).
"Deprotecting" refers to removing any protective groups present after the selective
reaction has been carried out.
6 "Disease" specifically includes any unhealthy condition of an animal or part thereof and
includes an unhealthy condition which may be caused by, or incident to, medical or veterinary
therapy applied to that animal, i.e., the "side effects" of such therapy.
g "Halo" means fluoro, chloro, bromo or iodo.
"Heteroalkylene" means alkylene, as defined above, wherein I to 5 of the carbon atoms
indicated is replaced by a heteroatom chosen from N, O or S (e.g., azaalkylene, oxaalkylene and
12 thiaalkylene, respectively), with the proviso that the oxygen, nitrogen and sulfur atoms contained
therein do not form bonds with other heteroatoms. For example, hetero(C3 ,2)alkylene is meant
to encompass aza(C3)alkylene which includes 3-azatrimethylene (-NHCH2CH2-),
15 2-azatrimethylene (-CH2 NH CH2-), etc.; ~-aza(C2 ~)alkylene which includes 2-azaethylene
(-NH CH2-), 3-azatrimethylene, 4-azatetramethylene (-NH CH2 CH2 CH2-) and
S-a~e~ ethylene (-NH-CH2-CH2-CH2-CH2-); oxa(C3)alkylene which includes as
18 3-oxatrimethylene (-O-CH2-CH2-), 2-oxatrimethylene (-CH2-0-CH2-), etc.; oxa(C5)alkylene such
as 3-oxapentamethylene (-CH2-CH2-0-CH2-CH2-), etc.; thia(C3)alkylene which includes
3-thiatrimethylene (-S-CH2-CH2-), 2-thiatrimethylene (-CH~-S-CH2-), etc.; ~I)-thia(C2 4)alkylene
21 which includes 2-thiaethylene (-NH-CH2-), 3-thiatrimethylene and 4-thiatetramethylene
(-S CH2 CH2 CH2-); di~7~(C6)alkylene which includes 2,5-diazahexamethylene
(-CH2-NH-CH2-CH2-NH-CH2-); azaoxa(C6)alkylene which includes 2,-oxa-5-azahexamethylene
24 (-CH2-0-CH2-CH~-NH-CH2-); and the like.
"Heteroarylene" means arylene, as defined above, wherein 1 to 5 of the carbon atoms
indicated are replaced by a heteroatom chosen from N, O or S (e.g., hetero(C5 6)arylene includes
27 furylene, thienylene, pyrrolylene, imidazolylene, pyridylene, etc.).
"Heterocycloalkylene" means cycloalkylene, as defined above, wherein I to 5 of the
carbon atoms indicated are replaced by a heteroatom chosen from N, O, or S
30 (e.g., hetero(C3 ,4)cycloalkylene includes 2,4-pyrrolidinylene, 2,4-pyrrolinylene,
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2,4-imidazolinylene, 2,4-imidazolinylene, 3,5-pyrazolinylene, 1,4-piperidylene,
1,4-piperazinylene, 2,5-quinuclidinylene, 2,5-morpholinylene, 1.3-isoindolinylene, etc.).
3 "Heterooxoalkylene" means alkylene, as defined above, wherein one of the number of
~ carbon atoms indicated is replaced by a heteroatom chosen from N, O or ~ and a carbon atom
adjacent to the heteroatom is replaced by a carbonyl group (C=O), e.g., azaoxoalkylene,
6 oxaoxoalkylene and thiaoxoalkylene, respectively, with the proviso that the oxygen, nitrogen and
sulfur atoms contained therein do not forrn bonds with other heteroatoms. For exarnple,
heterooxo(C4 6)alkylene is meant to encompass azaoxo(C3)alkylene which includes
9 2-aza-3-oxotrimethylene (-C(O) NH CH2-), 3-aza-2-oxotrimethylene (-NH C(O) CH2-), etc.;
oxaoxo(C~)alkylene which includes 2-oxa-3-oxotrimethylene (-C(O) O CH2-),
3-oxa-2-oxotrimethylene (-O C(O) CH2-), etc.; and thiaoxo(C3)alkylene which includes
12 2-thia-3-oxotrimethylene (-C-(O)-S-CH2-), 3-thia-2-oxotrimethylene (-S-C(O)-CH2-), etc.
"Leaving group" has the meaning conventionally associated with it in synthetic organic
chemistry, i.e., an atom or group displaceable under alkylating conditions, and includes, halogen,
hydroxy, alkylsulfonloxy (e.g., mesyloxy, ethanesulfonyloxy, etc.), arylsulfonyloxy
(e.g., benzenesulfonyloxy and tosyloxy, thienyloxy), dihalophosphinoyloxy,
tetrahalophosphaoxy, and the like.
18 "Optional" or "optionally" means that the subsequently described event or circumstance
may or may not occur, and that the description includes instances where the event or
circumstance occurs and instances in which it does not. For example, the phrase "optionally
21 substituted with one or more radicals" means that the group referred to may or may not be
substituted in order to fall within the scope of the invention.
"Ph~ ceutically acceptable N-Oxide" means compound in which nitrogens are in an
24 oxidized state (i.e., O-N) which are pharmaceutically acceptable, as defined below, and which
possess the desired pharmacological activity. The N-oxides of compounds of Formula I can be
prepared by methods known to those of ordinary skill in the art.
27 "Oxoalkylene" means alkylene, as defined above, wherein one of the number of carbon
atoms indicated is replaced by a carbonyl group (C=O), e.g., oxo(C3)alkylene includes
3-oxotrimethylene (-C(O) CH2 CH2-), etc
"Pathology" of a disease means the essential nature, causes and development of the
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disease as well as the structural and functional changes that result from the disease processes.
"Ph~ ceutically acceptable" means that which is useful in preparing a rh~ ceutical
3 composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable
and includes that which is acceptable for veterinary use as well as human pharmaceutical use.
"Pharmaceutically acceptable salts" means salts which are pharmaceutically acceptable,
6 as defined above, and which possess the desired pharmacological activity. Such salts include
acid addition salts formed with inorganic acids such as hydrobromic acid, hydrochloric acid,
nitric acid, phosphoric acid, sulfuric acid and the like; or with organic acids such as acetic acid,
9 bel~encsulfonic acid, benzoic acid, camphorsulfonic acid, p-chlorobenzene-sulfonic acid,
cinnamic acid, citric acid, cyclopentanepropionic acid, 1,2-ethanedisulfonic acid, ethanesulfonic
acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hexanoic acid,
12 heptanoic acid, o-(4-hydroxybenzoyl)benzoic acid, 2-hydroxyethanesulfonic acid,
hydroxynaphthoic acid, lactic acid, lauryl sulfuric acid, maleic acid, malic acid, malonic acid,
mandelic acid, methanesulfonic acid, 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid,
4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), muconic acid, 2-n~phth~lenesulfonic
acid, oxalic acid, 3-phenylpropionic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid,
succinic acid, tartaric acid, tertiary butylacetic acid, p-toluenesulfonic acid, trimethylacetic acid
18 and the like.
Pharmaceutically acceptable salts also include base addition salts which may be formed
when acidic protons present are capable of reacting with inorganic or organic bases. Acceptable
21 inorganic bases include alllmimlm hydroxide, calcium hydroxide, potassium hydroxide, sodium
carbonate and sodium hydroxide. Acceptable organic bases include diethanolamine,ethanolamine, N-methylglucamine, triethanolamine, tromethamine and the like.
24 "Phenylene" means the divalent aromatic radical -C6H4- and includes 1,4-phenylene,
1,3-phenylene and the like.
"Ph~ eutically acceptable prodrug derivatives" means derivatives of compounds of27 Formula I which are ph~ eutically acceptable, as defined above, and which are converted in
vivo to the corresponding non-derivatized forrn of a compound of Formula I. Such prodrugs
include compounds of Formula I which have N-acylated piperidyl (i.e., N(P)C5H9-), N-acylated
30 a7~1kylene (e.g., -N(P) CH2 CH2-), N-acylated amino (i.e., -NH~(P)), N-acylated amidino
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- I 3 -
(i.e.,-C(NP) NHP, -C(NH)-NHP or -C(NP)-NH2), N-acylated guanidino (i.e., -NHC(NP)-NHP,
-NH-C(NH)-NHP or -NH-C(NP)-NH2) groups, in which P is a group selected from -C(O)R'U,
3 wherein R'~ can be (C, ,0)alkyloxy or cis-2-(C, ,0)alkanoyloxyphenylvinyl,
3-(C~ )alkanoyloxybutyryl, R' I-X28-, wherein R~ ' is carboxy and X28 is (C"0)alkylene or
-C(O)-O-CH(R'2)-O-C(O)R' 3, wherein Rl2 is hydrogen, (C~ ~0)alkyl or (C3 ~O)cycloalkyl and R~ is
6 (C, ,0)alkyl.
"Protective group" has the meaning conventionally associated with it in synthetic organic
chemistry, i.e., a group which selectively blocks one reactive site in a multifunctional compound
9 such that a chemical reaction can be carried out selectively at another unprotected reactive site
and which can be readily removed after the selective reaction is completed.
"Protecting agent" means an agent which will react with a multifunctional compound and
12 create a protective group at a reactive site.
"Protected derivatives" in reference to a compound or a group means a derivative of
compound or group in which a reactive site or sites are blocked with protective groups.
5 Protected derivatives of compounds of Formula I are in themselves active as tryptase inhibitors
and are useful in the preparation of other compounds of Forrnula I. Suitable protecting groups
for reactive nitrogen atoms include tert-butoxycarbonyl, benzyloxycarbonyl and any other
18 suitable amino protective groups (e.g., see T W. Greene, Prolective Groups in Organic Synthesis,
John Wiley ~ Sons, Inc. I 98 l ).
"Symptomatology" of a disease means any morbid phenomenon or departure from the
21 norrnal in structure, function or sensation experienced by the patient and indicative of the
disease, their production and the indications they furnish.
"Therapeutically effective amount" means that amount which, when ;ltlmini~tered to an
24 animal for treating a disease, is sufficient to effect such treatment for the disease.
"Treating" or "treatment" of a disease includes preventing the disease from occurring in
an animal which may be predisposed to the disease but does not yet experience or display
27 symptoms of the disease, inhibiting the disease ( i.e., arresting its deve}opment) or relieving the
disease (i.e., c~ ing regression of the disease).
The term "q.s." means adding a ~uantity sufficient to achieve a stated function, e.g., to
30 bring a solution to the desired volume (i.e., 100%).
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The compounds of Formula I and the intermediates and staning materials used in their
ple~aLion are named in accordance IUPAC rules of nomenclature in which the characteristic
3 groups have decreasing priority for citation as the principle group as follows: acids, esters,
amides and amidines. Furthermore, for the purposes of this Application, when referring to a
divalent radical by written description the order of the number prefixes signifies the orientation
6 of its attachment. Similarly, when referring to a divalent radical by formula the way in which the
formula is presented signifies the orientation of ~ chment. For example, a compound of
Formula I in which R~ is 4-amidinobenzyl, Xl and Xg each are -NHC(O)-, x2 is
9 1,4-piperazinylene, X7 iS -C(O)O-, X~ iS 4,1-piperidylene and R2 is R9-X2~, wherein R9 is
piperid-4-yl and X2l is 3-azatrimethylene, is
illustrated by the following formula:
12 NH
H2N J~, ~ N ' X3-X4
/
N '~ f N - x7-x6
~N~N~
H O
which compound is named:
ci.s- 1 ,5-cyclooctylene 4-(4-amidinobenzylcarbamoyl)- 1 -piper~inec:~rboxylate
1~ 4-(2-piperid-4-ylaminoethylcarbamoyl)-1-piperidinecarboxylate when X3 and X7 each are
-C(O)O-, X4 and x6 each are a covalent bond, X5 iS cis-1,5-cyclooctylene and P is hydrogen;
3-{4-[2-(1-{cis-5-[4-(4-amidinobenzylcarbamoyl)piperazin-1 -ylcarbonyloxy]cyclooctyloxy-
1 8 carbonyl } piperid-4-ylcarbonylamino)ethylamino]piperid- 1 -ylcarbonyl } propionic acid when X3
and X7 each are -C(O~O-, X4 and x6 each are a covalent bond, X5 is cis- 1 ,5-cyclooctylene and P
is 3-carboxypropionyl;
21 4-[4-(4-amidinobenzylcarbamoyl)piperazin-1-ylcarbonyl~benzyl
4-(2-piperid-4-ylaminoethylcarbamoyl)-1-piperadinecarboxylate when X3 iS -C(O)-, X7 iS
-C(O)O-, X4 iS a covalent bond, x6 is methylene, X5 iS phenylene and P is hydrogen;
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_15_
1,4-tetramethylene 4-amidinobenzylcarbamoyl-1-piperazinecarboxylate when X3 and X7 are each
is -C(O)O- and X4-X5-X6 is 1,4-tetramethylene (i.e., -CH2-CH2-CH2-CH2-); and
3 N-4-amidinobenzyl-4- { 5- [4-(2-piperid-4-ylaminoethylcarbamoyl)piperid- 1 -ylcarbonyl]valeryl } -
- 1-piperazinecarboxamide when X3 and X7 are each is -C(O)- and X4-X5-X6 is 1,4-tetramethylene
(i.e., -CH2-CH2-CH2-CH2-) .
6 Presently Preferred Embodiments:
While the broadest definition of this invention is set forth in the Summary of the
Invention, certain compounds of Formula I are preferred. For example, preferred compounds of
g Formula I are those in which Xs is cis- 1 ,5-cyclooctylene and X4 and x6 each are a covalent bond,
X4-X5-X6 together are (C4 8)alkylene or X5 is 1,4-phenylene and X4 and x6 are (CO l)ethylene; X
and X9 are independently a covalent bond, -C(O)-, -NHC(O)-, -C(O)NH-, -N(CH3)C(O)- or
2 -S(0)2NH-, with the proviso that X~ and X9 are not both covalent bonds; X3 and X7 are
independently -C(O)- or -C(O)O-; x2 and x8 are independently -X~~-XI~-, wherein X~~ is a
covalent bond or methylene and X~ ~ is 4,1-piperidylene or 1 ,4-piperazinylene; R~ is R4-X'~- or
R5-X~3-, wherein R4 is amidino, guanidino or methylamino, X~2 is -Xl4-X~5-X~6-, wherein X~5 is
1,4-phenylene or 1,4-piperidylene, X'4 is (Cn,4)alkylene and Xl6 is (Cn~6)alkylene, wherein the
sum of nl4 and nl6 is 0, 1 or 2, R5 is piperid-4-yl and X~3 is (C2.3)alkylene; and R2 is R8-X20- or
8 R9-X2~-, wherein R8 is amino, amidino, guanidino, methylamino or l-iminoethyl, X20 is
-X22-X23-X24-, wherein X23 is ~rans-1,4-cyclohexylene, 1,4-phenylene, 4,1-pyridylene,
I ,4-piperidylene, X22 is (Cn22)alkylene and X24 is (Cn24)alkylene, wherein the sum of n22 and n24
21 is 1 or 2, R9 is benzoimidazol-5-yl, imidazol-l-yl, imidazol-4-yl, 2-imidazolin-2-yl,
4-methylimi(1~7ol-l -yl, 5-methylimidazol-1 -yl, 1-methylpiperid-4-yl, piperid-4-yl,
piperazin-l-yl, pyrid-3-yl, pyrid-4-yl, 1,4,5,6-tetrahydropyrimidin-5-yl or
24 1,4,5,6-tetrahydro-2-dioxopyrimidin-S-yl and X2' is (C, 6)alkylene, ~-aza(C2 s)alkylene,
3-oxotrimethylene, ~-thia(C2 4)alkylene, 3-oxo-2-azatrimethylene, 3-aza-2-oxotrimethylene or
-X25-X26-X2'-, wherein X26 is 1,4-phenylene, X25 is (Cn25)alkylene and X24 is (Cn27)alkylene,
- 27 wherein the sum of n25 and n27 is O or 1; and the pharmaceutically acceptable salts, ~-oxides,
prodrug derivatives and protected derivatives thereof.
More preferred compounds of Forrnula I are those in which X5 iS cis-l ,5-cyclooctylene
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-16-
and X4 and x6 each are a covalent bond or X4-Xs-X6 together are (C4 8)alkylene; X' and X9 are
independently a covalent bond -C(O)-, -NHC(O)-, -C(O)NH- or -S(0)2NH-, with the proviso that
3 X~ and X9 are not both covalent bonds; X3 and X7 are independently -C(O)- or -C(O)O-; x2 and
X3 are independently -X'~-X"-, wherein X'~ is a covalent bond or methylene and X" is
4,1-piperidylene or 1,4-piperazinylene; R~ is R4-X~2-, wherein R4 is amidino or guanidino and x~2
6 is -X~4-X~5-X~6-, wherein X~s is 1,4-phenylene or 1,4-piperidylene, X~4 is (Cn~4)alkylene and Xl6 is
(Cnl6)alkylene, wherein the sum of nl4 and nl 6isO,1 or 2; and R2 is R8-X20- or R9-X2l-, wherein
R8 is arnino or methylamino, X20 is -X22-X23-X24-, wherein X23 is trans-l ,4-cyclohexylene or
9 1,4-phenylene, X22 is (Cn22)alkylene and x~6 is (Cn24)alkylene, wherein the sum of n22 and n24 is
1 or 2, R9 is imidazol-l-yl, imidazol-4-yl, 4-methylimidazol-1-yl, 5-methylimidazol-1-yl,
piperid-4-yl or pyrid-4-yl and X2' is (C, 5)alkylene or 3-azatrimethylene; and the
2 pharrnaceutically acceptable salts, N-oxides, prodrug derivatives and protected derivatives
thereof.
Particularly preferred compounds of Formula I are those in which X5is
15 cis-1,5-cyclooctylene and X4 and X6 each are a covalent bond; X' and X9 are independently
-C~O)- or -NHC(O)-; X3 and X7 each are -C(O)O-; x2 and x8 are independently -X~~-XI '-,
wherein X'~ is a covalent bond and X" is 1,4-piperazinylene; R' is R4-X'2-, wherein R4 is
18 arnidino or guanidino and Xl2is -x'4-x'5-x'6-, wherein X~5isl,4-phenylene, X14is a covalent
bond and x~6 is methylene; and R2 is Rs-X20- or R9-X2l-, wherein R8 is amino, X20 is
-X22-X23-X24-, wherein X23 is trans- I ,4-cyclohexylene, XZ2 is a covalent bond and X24 is
21 methylene, R9 is piperid-4-yl and X2~ is ethylene or trimethylene; and the pharrnaceutically
acceptable salts, N-oxides, prodrug derivatives and protected derivatives thereof.
Particularly preferred compounds of Forrnula I are those in which X4-X5-X6 together are
24 (C4 8)alkylene; X~ and X9 are independently -C(O)- or -NHC(O)-; X3 and X7 are independently
-C(O)- or -C(O)O-; x2 and X~ each are -X~~-X~ ~-, wherein X~~ is a covalent bond and X~ ~ is
1,4-piperazinylene; R~ is R4-X~2-, wherein R4 is amidino or guanidino and Xl2 is -X~4-X~5-X'6-,
27 wherein X~5is 1,4-phenylene, X~4 is a covalent bond and Xl6 is methylene; and R2 is R~-X20-,
wherein R~ is amidino or guanidino and X20 is -X22-X23-X24-, wherein X23 is 1,4-phenylene, X22 is
a covalent bond and X24 is methylene; and the pharmaceutically acceptable salts, N-oxides,
30 prodrug derivatives and protected derivatives thereof.
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Most preferred compounds of Formula I are the following:
4-guanidinobenzyl 4-{7-[4-(4-guanidinobenzylcarbamoyl)piperazin-1-ylcarbonyl]-
3 heptanoyl}-l-piyel~inec~rboxamide;
4-guanidinobenzyl
4- { 8-[4-(4-guanidinobenzylcarbamoyl)piperazin- 1 -ylcarbonyl]octanoyl } -
6 l-piperazinecarboxamide;
4-guanidinobenzyl 4-{9-[4-(4-guanidinobenzylcarbamoyl)piperazin-1-ylcarbonyl]-
nonanoyl } -1 -piperazinecarboxamide;
9 4-amidinobenzyl
4-{7-[4-(4-amidinobenzylcarbamoyl)piperazin-1 -ylcarbonyl3heptanoyl}-
1 -piperazinecarboxamide;
12 cis-1,5-cyclooctylene 4-(4-amidinobenzylcarbamoyl)-1-piperazinecarboxylate
4-(2-piperid-4-ylethylcarbamoyl)- 1 -pip~ ecarboxylate;
1 ,5-pentamethylene di [4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylatel,
1 5 cis- 1 ,5-cyclooctylene 4-(4-amidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(4-piperid-4-ylbutyryl)- 1 -piperazinecarboxylate;
cis-1,5-cyclooctylene trans-4-(4-aminocyclohexylmethylcarbamoyl)-
18 l-piper~7:in~c~rboxylate 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate;
cis- 1 ,5-cyclooctylene 4-(4-amidinophenylacetyl)- 1 -piperazinecarboxylate
4-(4-piperid-4-ylbutyryl)- 1 -piperazinecarboxylate;
21 1,4-tetramethylene di[4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate];
cis- 1 ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(2-piperid-4-ylethylcarbamoyl)- 1 -piperazinecarboxylate;
24 4-guanidinobenzyl
4-{6-[4-(4-guanidinobenzylcarbamoyl)yipel~in-1-ylcarbonyl]hexanoyl}-
1 -piperazinecarboxamide;
27 cis- 1 ,S-cyclooctylene 4-(4-guanidinoberlzylcarbamoyl)- 1 -piperazinecarboxylate
- 4-(4-piperid-4-ylbutyryl)-1-piperazinecarboxylate;
cis-1,5-cyclooctylene 4-(4-guanidinophenylacetyl)-l-piper~7in~c~rboxylate
4-(2-piperid-4-ylethylcarbarnoyl)- 1 -yip~lA7i l.çc~rboxylate;
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cis- 1,5 -cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-(4-piperid-4-ylbutyryl)- 1 -piperazinecarboxylate;
3 4-guanidinobenzyl
4- { 5-[4-(4-guanidinobenzylcarbarnoyl)piperazin- 1 -ylcarbonyl]valeryl } -
1-piperazinecarboxamide;
6 3-oxa-1,5-pentarnethylene di[4-(4-guanidinophenylacetyl)piperazin-l-ylcarbonyl]; and
cis-1,5-cyclooctylene 4-(4-amidinophenylacetyl)-1-piper~inec~rboxylate
4-(2-piperid-4-ylethylcarbamoyl)-l-piperazinecarboxylate; and the pharmaceutically acceptable
g salts, N-oxides, prodrug derivatives and protected derivatives thereof.
Pharmacology and Utility:
The compounds of this invention are tryptase inhibitors. As such the compounds of
12 Forrnula I are useful for treating diseases, particularly immunomediated infl~mm~tory diseases in
which tryptase activity contributes to the pathology and/or symptomatology of the disease. For
example, immunomediated infl~mmzltory diseases in which tryptase activity contributes to its
15 pathology and/or symptomatology include asthrna, allergic rhinitis, rheumatoid spodylitis,
osteoarthritis, gouty arthritis, rheumatoid arthritis, arthritic conditions in general, urticaria,
angioedema, ec7em~tous dermatitis, anaphylaxis, hyper proliferative skin disease, peptic ulcers,
18 infl~mm~tory bowel disease, ocular and vernal conjunctivitis, infl~mm~tory skin conditions, and
the like.
Suitable in vi~ro assays for measuring tryptase activity and the inhibition thereof by
21 compounds are known (e.g., see Sturzebecher et al. (1992) Biol. Chem. Hoppe-Seyler
373:1025-1030). Typically, the assay will measures tryptase induced hydrolysis of peptide base
substrate. For further details of an in vitro assay for measurin~ tryptase activity see Example 33,
24 infra.
Suitable in vivo models of infl:~mm~tjon are known to those of ordinary skill in the art.
For exarnple, in vivo models for asthma are known (e.g., see Larsen (1991) Experimental Models
27 of Reversihle Airway Obstruction. In: West et al., eds. The J.ung: Scientific Foundations Raven
Press, New York). For further details of an in vitro model of astl~na see Example 2, infra.
Further, in vivo models of infl~mm~tory skin conditions (Walsh et al. (1995) Br. ~ P~armacol.
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114: 1343-1350; and Armstrong et al. (1995) Prostaglandins 49: 205-224), arthritic conditions
(Peacock et al. (1995) Cell Immunol. 160: 178-184; and Houri et al. (1995) Curr. Opin.
3 Pheumatol. 7: 201-205) and gastrointestinal diseases (Anthony et al. (1995) Int. J. Exp. Pathol.
- 7~: 215-224.; and Carter et al. ( 1995) Dig. Dis.Sci. 40: 192- 197) are known. For further details
of an in vivo assay for measuring asthmatic responses see Ex~mple 34, infra.
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Administration and Pharmaceutical Compositions:
In general, compounds of Formula I will be ~rlmini.~tered in therapeutically effective
3 amounts via any of the usual and acceptable modes known in the art, either singly or in
combination with another therapeutic agent. A therapeutically effective amount may vary widely
depending on the severity of the disease, the age and relative health of the subject, the potency of
6 the compound used and other factors. For example, therapeutically effective amounts of a
compound of Formula I for the treatment of asthma may range from 0. l micrograms per
kilogram body weight (~lg/kg) per day to 1 milligram per kilogram body weight (mglkg) per day,
g typically l ~g/kg/day to 0. l mglkglday. Therefore, a therapeutically effective amount for a 80 kg
~cthm~tic human patient may range from l 0 ,ug/day to l 0 mg/day, typically O. l mglday to
l 0 mglday.
12 Therapeutic agents that may be useful for :~tlmini~tration in combination with compounds
of Formula I in treating asthma include ~-adrenergic agonists (e.g., albuterol, terbutaline,
formoterol, fenoterol, prenaline and the like), methylxanthines (e.g., caffeine, theophylline,
aminophylline, theobromine and the like), cromoglycates (e.g., cromolyn, nedocromil, and the
like) and corticosteroids (e.g., beclomethasome, triamcinolone, flurisolide, dexamethasone and
the like). In general, one of ordinary skill in the art, acting in reliance upon personal knowledge
8 and the disclosure of this application, will be able to ascertain a therapeutically effective amount
of a compound of Formula I for treating a given infl~mm~tory disease.
The compounds of Formula I can be ~-lmini~tered as pharmaceutical compositions by one
21 ofthe following routes: oral, systemic (e.g., transdermal, intranasal or by suppository) or
parenteral (e.g., intramuscular, intravenous or subcutaneous). Compositions can take the form of
tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions,
24 suspensions, elixirs, aerosols, or any other appropriate composition and are comprised of, in
general, a compound of Formula I in combination with at least one pharrnaceutically acceptable
excipient. Acceptable excipients are non-toxic, aid ~lmini.~tration and do not adversely affect
27 the therapeutic benefit of the active ingredient. Such excipient may be any solid, liquid,
semisolid or, in the case of an aerosol composition, gaseous excipient that is generally available
to one of skill in the art.
Solid pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose,
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WO 98104537 PCT/US97/13422
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gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol
monostearate, sodium chloride, dried skim milk, and the like. Liquid and semisolid excipients
3 may be selected from water, ethanol, glycerol, propylene glycol and various oils, including those
- of petroleum, animal, vegetable or synthetic origin (e.g., peanut oil, soyhean oil? mineral oil,
sesarne oil, etc.). Preferred liquid carriers, particularly for injectable solutions, include water,
6 saline, aqueous dextrose and glycols.
Compressed gases may be used to disperse the active ingredient in aerosol form. Inert
gases suitable for this purpose are nitrogen, carbon dioxide, nitrous oxide, etc. Other suitable
g ph~ ceutical carriers and their formulations are described in A.R. Alfonso Remington's
Pharmaceutical Sciences 1985, 17th ed. Easton, Pa.: Mack Publishing Company.
The amount of a compound of Forrnula I in the composition may vary widely depending
12 upon the type of forrnulation, size of a unit dosage, kind of excipients and other factors known to
those of skill in the art of ph~ eutical sciences. In general, a composition of a compound of
Formula I for treating asthma will comprise from 0.01 %w to 1 0%w, preferably 0.3%w to I %w,
15 of active ingredient with the remainder being the excipient or excipients. Preferably the
ph~ ceutical composition is ~(lmini~tered in a single unit dosage form for continuous
treatment or in a single unit dosage form ad libitum when relief of symptoms is specifically
18 required. Representative ph~ ceutical formulations cont;~inin~ a compound of Formula I are
described in Example 34.
Chemistry:
2 1 The compounds of the invention are comprised of five distinct subunits (i.e., Rl-, -X2-,
-X4-X5-X6-, -X8- and R2-) which subunits are connected via carbonyl, formyloxy, amide,
sulfonamide, carbamate or urea linkages (i.e., -C(O)-, -C(O)O-, -OC(O)-, -C(o)N(R3)-,
24 -N(R3)C(o)-, -S(o)2N(R3)-, -N(R3)S(o)2-, -oC(o)N(R3)-, -N(R3)C(o)o-, -N(R3)C(o)N(R3)- or
-OC(O)O-). Methods for forming such linking groups are known and suitable reagents are
readily available (e.g., see, March"4dvanced Orgmic Chemistry, 4th Ed. (Wiley 1992); Larock,
27 Comprehensive Organic Transformations (VCH 1989); and Furniss et al., Vogel 's 7'extbook of
Practical Organic Chemistry, 5th Ed.. (Longman 1989).
The subunits comprising the compounds of Formula 1 can be assembled individually or as
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larger combinations of subunits. The following reaction schemes are representative methods for
pl~ lg compounds of Formula I. It is understood that the compounds of Formula I can be
3 prepared by other analogous procedures.
Compounds of Formula I in which x8 is 1 ,4-piperazinylene or 1 ,4-piperidylene and X9 iS
-C(O)-, -OC(O)- or -N~R3)C(o)- or in which x8 is (C~ 8)alkylene and X9 iS -C(o)N(R3)-,
6 -OC(O)N(R3)- or -N(R3)C(O)N(R3)- can be prepared by reacting a compound of Formula 1:
Rl-xl-x2-x3-x4\
y8 X7 X6 ~
or a protected derivative thereof, with a compound of the formula R2-Y9-C(O)~, or a protected
g derivative thereof, wherein L is a leaving group, Y9 is a bond, -O- or -N(R3)-, y8 iS
piperazin-1-yl, piperid-4-yl or HN(R3)-(C, ~)alkyl, respectively, and each R~, R2, R3, Xl, X2, X3,
X4, Xs, x6 and X7 are as defined in the Summary of the Invention, and then deprotecting when
t 2 neces~ry. Alternatively, compounds of Formula I in which x8 is 1 ,4-piperazinylene or
I ,4-piperidylene and X9 is -NHC(O)- or in which x8 is (C~ 8)alkylene and X9 is -NHC(O)N(R3)-
can be prc;paled by reacting an ~lol)~iate compound of Formula l, or a protected derivative
15 thereof, with an isocyanate of the formula R2-NC(O), or a protected derivative thereof, and then
deprotecting when neces~y (for further details see Exarnple 8, infra.).
In an analogous fashion, compounds of Formula I in which x2 is 1,4-piperazinylene or
8 1,4-piperidylene and Xl is -C(O)-, -OC(O)- or -N(R3)C(o)- or in which X2is (C, 8)alkylene and
Xl is -C(o)N(R3)-, -oC(o)N(R3)- or -N(R3)C(O)N(R3)- can be prepared by reacting a compound
of Formula 2:
21 y2 X3 x4
R2 Xg X8 X7 Xfi ~
. ..
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or a protected derivative thereof, with a compound of the forrnula R'-Y'-C(O)L, or a protected
derivative thereof, wherein L is a leaving group, yl is a bond, -O- or -N(R3)-, y2 iS
3 piperazin-1-yl, piperid-4-yl or HN(R3)-(CI 8)allcyl, respectively, and each R~, R2, R3, X3, X4, X5,
X6, X7, X8 and X9 are as defined in the Summary of the Invention, and then deprotecting when
necessary. Alternatively, compounds of Formula I in which x2 is 1,4-piperazinylene or
6 1 ,4-piperidylene and X~ is -NHC(O)- or in which x2 is (Cl 8)alkylene and X~ is -NHC(O)N(R3)-
can be prepared by reacting a compound of Forrnula 2, or a protected derivative thereof, with an
isocyanate of the forrnula R'-NC(O), or a protected derivative thereof, and then deprotecting
9 when necessary (for further details see Example 14(b), infra.).
Compounds of Formula I in which R' equals R2; X2 and/or x8 is 1,4-piperazinylene or
1,4-piperidylene; X~ is -C(O)-, -OC(O)- or -N(R3)C(o)-; and X9 iS -C(O)-, -OC(O)- or
2 -N(R3)C(O)- and/or in which X2 and/or x8 is (C~ 8)alkylene; X~ is -C(O)N(R3)-, -oC(o)N(R3)- or
-N(R3)C(O)N(R3)-; and X9 -C(o)N(R3)-, -oC(o)N(R3)- or -N(R3)C(o)N(R3)- can be prepared by
reacting a compound of Formula 3:
y8 X7X6 ~
or a protected derivative thereof, with 2 or more molar equivalents of a compound of the formula
R~-Y'-C(O)L, or a protected derivative thereof, wherein L is a leaving group, Y' is a bond, -O- or
8 -N(R3)-, y2 and y8 are independently 1~ip~ -1-yl, piperid-4-yl or HN(R3)-(C~ 8)alkyl and each
R~, R3, X3, X4, X5, X6 and X7 are as defined in the Surnmary of the Invention, and then
deprotecting when necessary. Alternatively, compounds of Forrnula I in which R' equals R2; X2
21 and/or x8 is 1,4-piperazinylene or 1,4-piperidylene; X' is -NHC(O)- and/or X9 iS -NHC(O)-
and/or in which x2 and/or x8 is (Cl 8)alkylene and X~ is -NHC(o)N(R3)- and/or X9 is
-NHC(O)N(R3)- can be prepared by reacting a compound of Formula 3, or a protected derivative
24 thereof, with two or more molar equivalents of an isocyanate of the formula R~-NC(O), or a
protected derivative thereof, and then deprotecting when necessary (for further details see
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-24-
Example 10, infra.).
Compounds of Formula I in which X' is -N(R3)C(o)-, -N(R3)C(o)O- or
3 -N(R3)C(o)N(R3)- can be prepared by reacting an amine of the formula R'-N(R3)H, or a
protected derivative thereof, with a compound of Formula 4:
LC(o)-YI-X2-X3-Xt
R2 Xg x8 x7 x6 ~
6 or a protected derivative, wherein L is a leaving group, Y' is a bond, -O- or -N(R3)- and each R',
R2, R3, X2, X3, X4,X5, X6, X7, X8 and X9 are as defined in the Summary of the Invention (for
further details see Example 20, infra.).
9 Compounds of Formula I in which X2is1,4-piperazinylene or 4,1-piperidylene and X3is
-C(O)-, -C(O)O- or -C(O)N(R3)- or in which x2 is (C, 8)alkylene and X3 is -N(R3)C(O)-,
-N(R3)C(O)o- or -N(R3)C(O)N(R3)- can be prepared by reacting a compound of the
12 forrnula R'-X'-Y2, or a protected derivative thereof, with a compound of Formula 5:
LC(o)--'3-X4
R2XgX8X7
or a protected derivative thereof, wherein L is a }eaving group, Y3iS a bond, -O- or -N(R3)-, Y2is
piperazin-1-yl, piperid-4-yl or HN(R3)-(C, 8)alkyl, respectively, and each R', R2, R3, X', X2, X3,
X4,X5, X6, X7, x8 and X9 are as defined in the Summary of the Invention (for further details see
Example 31, infra.).
8 Compounds of Formula I in which X2 and X8 each are 1,4-piperazinylene or
4,1-piperidylene and X3 and X7 are independently -C(O)-, -C(O)O- or -C(O)N(R3)- or in which
X2 and X8each are (C, 8)alkylene or hetero(C, 8)alkylene and X3 and X7 are independently
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-25-
-N(R3)C(o)-, -N(R3)C(o)o- or -N(R3)C(O)N(R3)- can be ~l~pal~d by reacting two or more
molar equivalents of a compound of the formula R'-X'-Y2, or a protected derivative thereof~ with
3 a compound of Formula 6:
LC(O) Y3-X4
LC(o)Y7-X6
or a protected derivative thereof, wherein L is a leaving group, Y3 and Y7 are independently a
6 bond, -O- or -N(R3)-, y2iS piperazin-l-yl, piperid-4-yL HN(R3)-(C,.8)alkyl orHN(R3)-hetero(CI 8)alkyl, respectively, and each R~, X~, X4, X~ and x6 are as defined in the
Summary of the Invention (for further details see Example 32, infra.).
g The acylation reactions described above can be carried out by reacting together an
activated ester (e.g., an acid chloride derivative) and an appropriate nucleophile in the presence
of a suitable organic base (e.g., N,~T-diisopropylethylamine (DIEA), N-methylmorpholine, etc.
12 preferably DIEA) and suitable solvent (e.g., N,N-dimethylforrnamide (DMF), tetrahydrofuran
(THF), dichloromethane, etc.) at 20 to 30~C, typically at approximately 23 ~C, for several
minutes to 24 hours. Alternatively, the acylation can be effected by reacting together an
15 applol)l,ate carboxylic acid and nucleophile in the presence of a suitable coupling reagent (e.g.,
1-~3-dimethylarninopropyl)-3-ethylcarbodiimide, etc.) and a suitable solvent (e.g., DMF, etc.) at
20 to 30~C, typically at approximately 23 ~C, for several hours to several days. The reactions
8 described above for the plel)aldlion of compounds of Formula I and the conditions for effecting
the reactions are illustrative and one of ordinary skill in the art will recognize that other reaction
conditions can be applied and different starting materials can be used to prepare the compounds
21 ofthe invention.
Deprotection can be effected by any means which removes the protective group and gives
the desired product in reasonable yield. A detailed description of the techni~ues applicable to the
24 creation of protective groups and their removal can be found in T.W. Greene, Protective Groups
in Organic Synthesis, John Wiley & Sons, Inc. 1981.
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-26-
Generally, the starting materials useful in preparing the compounds of Formula I and the
intermediates useful in p,epa,illg the eompounds of Formula I are commereially available or can
3 be readily prepared by those of ordinary skill in the art. For example, intermediates use~ul in
preparing the compounds of Formula I are conveniently prepared by the aeylation reactions
described above. When necessary suitable protection chemistry is employed to direct the
6 reaetion to the desired reactive site when multiple reactive sites are present in the starting
materials.
A convenient starting material for preparing eompounds of Formula I in which X3 and X'
9 each are -C(O)O- is a eompound of Formula 7:
LC(O)-O-~
LC(O)-O-X
in whieh eaeh X4,X5 and x6 are as defined in the Summary of the Invention. For example,
12 eompounds of Formula 6 in which L is chloro can be prepared by reacting a corresponding diol
(e.g., cis-l,S-cyclooct~ns~liol, trans-1,4-cyelohexylendimethanol, 1,4-phenylenedimethanol, etc.)
with triphosgene (for further details see Example 5, infra.).
Intermediates useful in pl~p~hlg compounds of Formula I in which such intermediate
contains a amidino group can be prepared by treating a eorresponding nitrile with hydrogen
ehloride in ethanol and then reaeting with ammonia.
18 Additional Proeesses for Preparing Compounds of Formula I:
Compounds of Forrnula I in which R4 is guanidino can be prepared by reaeting a
eorresponding compound of Formula I in which R4 is amino with cyanarnide. The reaction is
21 earried out by treating the amine with hydrogen ehloride and then reaeting neat with an excess of
cy~n~micle at approximately 65 ~C for about two hours (for further details see Example 15,
infra.).
24 Compounds of Formula I may be prepared as ph~ eeutieally acceptable aeid addition
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wo g8/04537 rCT/US97113422
salts by reacting the free base forrns of a compound of Formula I with a pharrnaceutically
acceptable inorganic or organic acid. Alternatively, the ph~ reutically acceptable base
3 addition salts of compounds of Forrnula I may be prepared by reacting the free acid forms of
compounds of Formula I with pharrnaceutically acceptable inorganic or organic bases. Inorganic
and organic acids and bases suitable for the preparation of the pharmaceutically acceptable salts
6 of compounds of Formula I are set forth in the definitions section of this application.
Alternatively, the salt forrns of the compounds of Formula I may be prepared using salts of the
starting materials or intermediates.
g The free acid or free base forms of the compounds of Formula I can be prepared from the
corresponding base addition salt or acid addition salt form. For example, compounds of Formula
I in an acid addition salt form may be converted to the corresponding free base by treating with a
12 suitable base (e.g., ammonium hydroxide solution, sodium hydroxide, etc.). Compounds of
Formula I in a base addition salt form may be converted to the corresponding free acid by
treating with a suitable acid (e.g., hydrochloric acid, etc).
The N-oxides of compounds of Formula I can be prepared by methods known to those of
oldilla~y skill in the art. For exarnple, N-oxides can be prepared by treating an unoxidized forrn
of the compound of Formula I with an oxidizing agent (e.g., trifluoroperacetic acid, permaleic
18 acid, perbenzoic acid, peracetic acid, meta-chloroperoxybenzoic acid, etc.) in a suitable inert
organic solvent (e.g., a halogenated hydrocarbon such as methylene chloride) at approximately
0~C. Alternatively, the N-oxides of the compounds of Formula I can be prepared from the
21 N-oxide of an a~ropliate starting m~qteri~l.
Compounds of Forrnula I in unoxidized form can be prepared from N-oxides of
compounds of Formula I by treating with a reducing agent (e.g., sulfur, sulfur dioxide, triphenyl
24 phosphine, lithium borohydride, sodium borohydride, phosphorus trichloride, tribromide, etc.) in
an suitable inert organic solvent (e.g., acetonitrile, ethanol, a~ueous dioxane, etc.) at 0 to 80~C.
Prodrug derivatives of the compounds of Formula I can be prepared by methods known to
27 those of ordinary skill in the art (e.g., for further details see Saulnier et al.(l994), Bioorganic and
Medicinal Chemistry Letters. 4:198~). For example, ~ p.;ate prodrugs can be prepared by
reacting a non-derivatized compound of Formula I with a suitable cal~alllylating agent
30 (e.g., I,1-acyloxyalkylcarbonochloridate,para-nitrophenyl carbonate, etc.).
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Protected derivatives of the compounds of Formula I can be made by means known to
those of ordinary skill in the art. A detailed description of the techni~ues applicable to the
3 creation of protective groups and their removal can be found in T.W. Greene, Protective Groups
in Organic Synthesis, John Wiley & Sons, Inc. 1981.
In summary, an aspect of this invention is a process for plepa~ g compounds of
6 Formula I, which process comprises:
(a) reacting a compound of Formula 1:
Rl-XI-X2-X3-X4
y8X7 x6
g or a protected derivative thereof, with a compound of the forrnula R2-Y9-C(O)L, or a protected
derivative thereof, wherein L is a leaving group, Y9iS a bond, -O- or -N(R3)-, Y8iS
piperazin-1-yl, piperid-4-yl or HN(R3)-(CI 8)alkyl, respectively, and each R~, R2, R3, X', X2, X3,
12 X4, Xs, X6 and X7 are as defined in the Surnmary of the Invention, and then deprotecting when
necessary, to give a compound of Formula I, in which x8 is 1,4-piperazinylene or1,4-piperidylene and X9 is -C(O)-, -OC(O)- or -N(R3)C(o)- or in which x8 is (C,.8)alkylene and
X9is -C(O)N(R3)-, -OC(O)N(R3)- or -N(R3)C(O)N(R3)-;
(b) reacting a compound of Formula 1, or a protected derivative thereof, with an isocyanate
of the formula R7-NC(O), or a protected derivative thereof, and then deprotecting when
8 necessary, to give a compound of Formula I in which X8isI,4-piperazinylene or
1,4-piperidylene and X9is -NHC(O)- or in which x8 is (C, 8)alkylene and X9 is -NHC(O)N(R3)-;
(c) reacting a compound of Formula 2:
21 y2 x3 X4
R2X9X8X7x6~
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WO 98104537 PCT/US97/13422
-29-
or a protected derivative thereof, with a compound of the formula R'-Y'-C(O)L, or a protected
derivative thereof, wherein L is a leaving group, Y' is a bond, -O- or -N(R3)-, Y2iS
3 piperazin-l-yl, piperid-4-yl or HN(R3)-(C, 8)alkyl, respectively, and each R~, R2, R3, X3, X4, X5,
X6, X7,X8 and X9 are as defined in the Summary of the Invention, and then deprotecting when
~Pcess~ry, to give a compound of Formula I in which x2 is 1 ,4-pip.,ld~irlylene or
6 1 ,4-piperidylene and X~ is -C(O)-, -OC(O)- or -N(R3)C(o)- or in which x2 is (C,.8)alkylene and
Xl is -C(o)N(R3)-, -oC(o)N(R3)- or-N(R3)C(O)N(R3)-,
(d) reacting a compound of Formula 2, or a protected derivative thereof, with an isocyanate
9 of the formula Rl-NC(O), or a protected derivative thereof, and then deprotecting when
n~ces.c~ry, to give a compound of Formula I in which x2 is 1,4-piperazinylene or1,4-piperidylene and X~ is -NHC(O)- or in which X2is (C~ 8)alkylene and X' is -NHC(o)N(R3)-;
12 (e) reacting a compound of Forrnula 3:
y2 x3 x4\
y8X7 x6
or a protected derivative thereof, with 2 or more molar equivalents of a compound of the formula
5 R'-Y'-C(O)L, or a protected derivative thereof, wherein L is a leaving group, Y~ is a bond, -O- or
-N(R3)-, y2 and y8 are independently piperazin- 1 -yl, piperid-4-yl or HN(R3)-(C, 8)alkyl and each
R~, R3, X3, X4, Xs, X6 and X7 are as defined in the Summary of the Invention, and then
1~ deprotecting when necess~ry, to give a compound of Forrnula I in which R~ equals R2; X2 and/or
X8 is 1,4-pil)eld~h,ylene or 1,4-piperidylene; X~ is -C(O)-, -OC(O)- or -N(R3)C(O)-; and X9 is
-C(O)-, -OC(O)- or -N(R3)C(o)- and/or in which x2 and/or x8 is (Cl 8)alkylene; X~ is
2 1 -C(O)N(R3)-, -oC(o)N(R3)- or -N(R3)C(o)N(R3)-; and X9 -C(O)N(R3)-, -oC(o)N(R3)- or
-N(R3)c(o)N(R3)-;
(f) reacting a compound of Formula 3, or a protected derivative thereof, with two or more
24 molar equivalents of an isocyanate of the forrnula R'-NC(O), or a protected derivative thereof,
and then deprotecting when nrce~S~ry, to give a compound of Formula I in which R~ equals R2;
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X2 andlor x8 is 1 ,4-piperazinylene or 1 ,4-piperidylene; X' is -NHC~O)- and/or X9 is -NHC(O)-
andlor in which x2 and/or x8 is (Cl 8)alkylene and X~ is -NHC(o)N(R3)- and/or X9 is
3 -NHC(O)N(R3)-;
(g) reacting an amine of the forrnula R'-N(R3)H, or a protected derivative thereof, with a
compound Forrnula 4:
6 LC(o)-yl-x2-x3-x~ X5
R2XgX~X7X6~
or a protected derivative thereof, wherein L is a leaving group, Y' is a bond, -O- or -N(R3)- and
each Rl, R2, R3, X2,X3,X4, X5,X6, X7,X8 and X9 are as defined in the Summary of the Invention,
g and then deprotecting when necessary, to give a compound of Formula I in which Xl is
-N(R3)C(O)-, -N(R3)C(o)o- or -N(R3)C(O)N(R3)-;
(h) reacting a compound of the formula R'-X'-Y2, or a protected derivative thereof, with a
12 compound of Formula 5:
LC(o)-Y3-X~
R~-X9 X8 X7 X6
or a protected derivative thereof, wherein L is a leaving group, Y3 is a bond, -O- or -N(R3)-, Y2iS
15 pip~in-1-yl, piperid-4-yl or HN(R3)-(C~ 8)alkyl, lcspe~ ely, and each R~, R2, R3, X~, X2, X3,
X4,Xs, X6,X7, x8 and X9 are as defined in the Summary of the Invention, and then deprotecting
when necessary, to give a compound of Formula I in which x2 is 1,4-piperazinylene or
18 4,1-piperidylene and X3 is -C(O)-, -C(O)O- or -C(o)N(R3)- or in which X2iS (C1 8)alkylene and
X3 is -N(R3)C(O)-, -N(R3)C(O)O- or -N(R3)C(o)N(R3)-;
(i) reacting 2 or more molar equivalents of compound of the formula R'-X'-Y2, or a
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protected derivative thereof, with a compound of Formula 6:
LC(o)-Y3-X4
LC(o)Y7-X
3 or a protected derivative thereof, wherein L is a leaving group, Y3 and Y7 are independently a
bond, -O- or -N(R3)-, Y2iS piperazin-l-yl, piperid-4-yl, HN(R3)-(C, 8)alkyl or
HN(R3)-hetero(C, 8)alkyl and each R', X', X4, X5 and x6 are as defined in the Summary of the
6 Invention, and then deprotecting when necessary, to give a compound of Formula I in which x2
and X8 each are 1 ,4-piperazinylene or 4,1-piperidylene and X3 and X7 are independently -C(O)-,
-C(O)O- or -C(O)N(R3)- or in which x2 and x8 each are (C, ~)alkylene or hetero(C, 8)alkylene
9 and X3 and x2 are independently -N(R3)C(O)-, -N(R3)C(o)o- or -N(R3)C(o)N(R3)-, respectively;
(j) optionally reacting a compound of Formula I in which R4 is amino with cyanamide to
give a compound of Formula I in which R4 is guanidino,
2 (k) optionally further converting a compound of Formula I into a pharmaceutically
acceptable salt;
(l) optionally further converting a salt form of a compound of Formula I to non-salt form;
5 (m) optionally further converting an unoxidized form of a compound of Formula I into a
ph~rrn~celltically acceptable N-oxide;
(n) optionally further an N-oxide form of a compound of Formula I its unoxidized form;
18 (o) optionally further converting a non-derivatized compound of Formula I into a
pharrnaceutically prodrug derivative; and
(p) optionally further converting a prodrug derivative of a compound of Forrnula I to its
21 non-derivatized form.
In any of the above processes, a reference to Formula I refers to such Formula wherein
- each R', R2, R3, X', X2, X3, X4, Xs,X6, X7 X8 and X~ are as defined in their broadest definitions
24 set forth in the Summary of the Invention, with the processes applying particularly well to the
presently preferred embodiments.
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-32-
Examples:
EXAMPLE 1
3 tert-Butyl 4-arninobenzylcarbamate hydrochloride
4-Aminobenzylarnine (50.34 g, 0.412 mol) in dichloromethane (200 mL) was placed in a
one liter 3 neck round bottom flask fitted with a mechanical stirring a~ ald~us and the solution
6 was cooled to 0~C. di-tert-Butyl dicarbonate (89.9 g, 0.412 mol.) in dichloromethane (200 mL)
was added dropwise to the solution over 30 minutes and the resulting suspension was stirred
2 hours at 0~C giving a nearly homogeneous solution. The dichloromethane solution
9 subsequently was washed with aqueous sodium hydroxide (1.0 M1 500 mL) and then water
(500 mL). The organic layer was dried (MgSO4), filtered and concentrated in vac2~o giving a
yellow oil. The oil was taken into ethyl ether: methanol (2:1, 225 mL) and the solution was
12 cooled to 0~C, acidifled with hydrogen chloride in dioxane (4.0 M, 115 mL, 0.412 mol.) and
combined with ethyl ether (200 mL) giving a thick pale yellow precipitate. The precipitate was
collected by filtration and washed with additional ethyl ether (500 mL). Drying in vacllo gave
15 tert-butyl 4-aminobenzylcarbamate hydrochloride (100.23 g, 0.387 mol, 94% yield) as a pale
yellow solid; 'H-NMR (300MHz, DMSO-d6): 10.40-10.20 (br s, 3H), 7.40 (tr, lH), 7.30 (s, 4H),
4.10 (d, 2H), 1.40 (s, 9H).
18 EXAMPLE 2
tert-Butyl 4-guanidinobenzylcarbamate
Cyanamide (100 g, 2.4 mol) was placed in a 500 mL round bottom flask and heated to
21 between 60 and 65~C until the material completely melted and then tert-butyl
4-arninobenzylcarbamate hydrochloride (25.3 g, 97.8 mmol.), prepared as in Example 1, was
added directly to the liquid cyanamide giving a yellow solution. The solution was stirred 2 hours
24 at between 60 and 65 ~C and then water (100 mL) was added. The aqueous mixture was cooled
to room ~ ule and washed with ethyl ether (1 L). The organic phase was back extracted
with water (2x, 100 mL) and the combined aqueous layers were washed with ethyl ether (500
. .
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W O 98/04537 PCTrUS97/13422
mL), cooled in an ice water bath and then basified with aqueous sodium hydroxide (10 M,
100 mL) giving an insoluble oil which slowly cryst~ e~l The crystals were collected by
3 filtration washed with water. Drying in vacuo gave tert-butyl 4-guanidinobenzylcarbamate (18.3
g, 69.24 mmol, 70.8% yield) as a colorless crystalline solid; lH-NMR (300MHz,
DMSO-d6): 9.70 (s, lH), 7.42 (tr, lH), 7.40 (s, 4H), 7.25 (d, 2H), 7.15 (d, 2H), 4.10 (d, 2H), 1.40
6 (s, 9H).
EXAMPLE 3
tert-Butyl 4-chlorocarbonyl-1-piperazinecarboxylate
9 Triphosgene (25 g, 84.2 mmol) was taken into dichloromethane ~200 mL) and the
resulting solution cooled to 0~C. A mixture of tert-butyl l-piperazinecarboxylate (40 g,
214.~ mmol) and pyridine (35 mL, 432.7 mmol) in dichloromethane (100 mL) then was added
2 dropwise to the triphosgene solution and the reaction mixture was allowed to warm to room
temperature over 30 minutes. The mixture was quenched with aqueous hydrochloric acid (O.lN,
200 mL) and the aqueous phase was washed with dichloromethane (50 mL). The combined
organic layers were dried (MgSO4) and filtered. Concentrating in vacuo gave tert-butyl
4-chlorocarbonyl-1-piperazinecarboxylate (45.6 g, 71.6 mmol, 85% yield) as a yellow solid;
~H-NMR (300MHz, CDCI3): 3.70 (m, 2H), 3.60 (m, 2H), 3.50 (m, 4H), 1.50 (s, 9H).
18 EXAMPLE 4
tert-Butyl 4-(4-guanidinobenzylcarbamoyl)-1-piper~7inec~rboxylate trifluoroacetate
tert-Butyl 4-guanidinobenzylcarbamate (41.77 g, 0.158 mol.), prepared as in Example 2,
21 was treated with trifluoroacetic acid (TFA) (100 mL) for 30 minutes at room temperature. The
resulting nearly colorless liquid was concentrated in vacuo at 45 ~C and the residue was triturated
with ethyl ether (3x, 400 mL) and dried in vacuo to a colorless foam. The residue was dissolved
24 in methanol (200 mL) and then DIEA (55 mL, 0.32 mol, amount based on estimated excess TFA
present) was added to the solution. The mixture was cooled to 0~C and then tert-butyl
4-chlorocarbonyl-1-pi~,cl~inecarboxylate (39.3 g, 0.158 mol.), prepared as in Example 3, in
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dichloromethane (120 mL) was added. An additional amount of DIEA (30 mL) was added and
the reaction mixture was allowed to warm to room temperature, stirred 12 hours and concentrated
3 in vacuo giving an orange oil. The oil was combined with water (200 mL) giving a thick
pl~i~ le. Recryst~ tion of the precipitate from acetonitrile and ether gave tert-butyl
4-(4-guanidinobenzylcarbamoyl)~ )eld~inecarboxylate trifluoroacetate (62.0 g, 0.126 mol,
6 80% yield) as a pale yellow solid; 'H-NMR (300MHz, DMSO-d~): 10.15 (s, 1 H), 9.10 (br s, 2H),
7.65 (s, 4H), 7.40 (tr, lH), 7.25 (dd AB, 4H), 4.25 (d, 2H), 3.55 (m, 4H), 3.10 (s, 4H),
Electrospray LRMS: Calculated for Cl3H20N6O: MH+: 277.4; MH2+212: 139.2,
g Found: MH+: 277.4; MH2+2/2: 139.3.
.. . . .
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EXAMPLE 5
cis-1,5-Cyclooctylene di(chloroformate)
- 3 cis-1,5-Cyclooctanediol (20.2 g, 0.14 mol.) was taken into acetonitrile (250 mL) and
potassium carbonate (41.4 g, 0.3 mol.) was added to the mixture giving a suspension. The
suspension was cooled to 0~C under a nitrogen atmosphere and then phos~ene (1.9M in toluene,
6 220 mL, 0.42 mol.) was added dropwise over one hour. The suspension was warmed to room
temperature and stirred 12 hours and then ether (1 L) was added. The suspension was filtered
free of insoluble salts and concentrated. Recrys~11i7~tion of the residue from hexane gave
9 cis- 1 ,5-cyclooctylene di(chloroformate) as a colorless crystalline solid. Further purification can
be effected with silica gel flash chromatography using hexane:ethyl ether (10:1) as eluent;
'H-NMR (300MHz, CDC13): 5.00-4.85 (m, 2H), 2.20-1.60 (m, 12H).
12 EXAMPLE 6
cis-1,5-Cyclooctylene chloroformate 4-tert-butoxycarbonyl-1-piperazinecarboxylate
cis-1,5-Cyclooctylene di(chloroformate) (1.91 g, 7.1 mmol), prepared as in Example 5, in
dichloromethane (25 mL) was added dropwise to a mixture of tert-butyl l-piperazinecarboxylate
(1.3 g, 7.1 mmol) and DIEA (1.3 mL, 7.1 mmol) in dichloromethane (25 mL). The mixture was
stirred 15 minutes at room temperature and then a workup with 0.1 M aqueous hydrochloric acid
8 was performed. The dichloromethane layer was dried (MgSO4), filtered and concentrated.
Purifying from the residue by silica gel flash chromatography using ethyl ether and hexanes as
eluent gave cis-1,5-cyclooctylene chloroformate 4-tert-butoxycarbonyl-1-piperazinecarboxylate
21 (660 mg, 1.6 mmol, 22% yield) as a colorless oil; ~H-NMR (300MHz, CDCI3): 5.00-4.90 (m?
lH), 4.80-4.70 (m, lH), 3.40 (s, 8H), 2.05-1.40 (m, 12), 1.40 (s, 9H).
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EXAMPLE 7
cis-l,S-Cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
34-tert-butoxycarbonyl- 1 -piperazinecarboxylate
tert-Butyl 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate trifluoroacetate
(383.7 mg, 1.06 mmol), prepared as in Example 4, was treated with trifluoroacetic acid (1 mL)
6 neat for 10 minutes at room temperature. The mixture was concelltl~d in ~Jac~o giving a
colorless oil. The oil was then taken into water (15 mL) and the pH of the aqueous solution was
adjusted to between 7 and 8 with 5M aqueous sodium hydroxide added dropwise.
g cis-l,S-Cyclooctylene chloroforrnate 4-tert-butoxycarbonyl-1-piperazinecarboxylate (444.7 mg,
1.06 mmol), prepared as in Example 6, in THF (10 mL) was added to the aqueous solution and
the pH was continually adjusted with lM aqueous sodium hydroxide added dropwise until no
12 further change in pH was observed. The mixture was concentrated in vacuo removing the bulk
of THF and then ethyl ether (5 mL) and SM aqueous sodium hydroxide (sufficient to adjust the
pH to 14) was added giving a thicic white suspension. The suspension was allowed to stand for
5 15 to 30 minutes at room temperature and then the precipitate was collected by filtering and
washed with water (2x, l S mL). Drying in vacuo gave cis- l ,S-cyclooctylene
4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate 4-tert-butoxycarbonyl-
8 l-pi~eld2i~lecarboxylate (527 mg, 0.82 mmol, 77% yield) as a colorless solid; 'H-NMR
(300MHz, DMSO-d6): 7.05 (d, 2H), 7.00 (tr, lH), 6.70 (d, 2H), S.10 (br, 3H), 4.65 (m, 2H), 4.15
(d, 2H), 3.30 (s, 16H), 1.90-1.40 (m, 12H), 1.40 (s, 9H).
21EXAMPLE 8
cis- 1 ,5-Cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate trifluoroacetate
24(Compound 1)
The following is the prel)al~lion of a compound of Formula I in which Rl is
27 4-guanidinobenzyl, R2 is 2-piperid-4-ylethyl, X' and X9 each are -NHC(O)-, x2 and x8 each are
1,4-pip~d~illylene,X3 and X7 each are -C(O)O-, X4 and x6 each are a covalent bond and X~ is
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cis- 1,5-cyclooctylene.
3 cis- 1,5-Cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-tert-butoxycarbonyl-1-piperazinecarboxylate (818 mg, 1.24 mmol.), p~aled as in Example 7,
was treated with TFA (2 mL) neat for 10 minutes. The mixture was concentration in vacuo
6 giving a colorless oil. The residue was triturated with ethyl ether (2x, 10 mL) and dried in vacuo
giving a colorless foam. The residue was then taken into DMF (2 mL) and then DIEA (700 mL,
4.0 mmol.) and tert-butyl 4-(2-isocyanatoethyl)-1-piperidinecarboxylate (3.2 mL, 0.39 M in
g DMF, 1.25 mmol.) were added. The mixture was stirred 12 hours and then concentrated in
vacuo. The residue was triturated with water (2x, 5 mL) and dried in vacuo giving a yellow
solid. The solid was then treated with TFA (2 mL) and the mixture was concentrated in vacuo.
12 The residue was taken into water. Purifying from the aqueous mixture by ~lepaldlive reverse
phase HPLC ~ave cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-
1-piperazinecarboxylate 4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate as an
amorphous colorless solid; Plasma Desorption LRMS: Calculated for C3sH55NI0o6 MH+: 712.9,
Found: MH+: 713.2.
Proceeding as in Exarnple 8 and substituting different starting materials the following
18 compounds of Forrnula I were prepared:
cis- l ,5-cyclooctylene 4-(4-aminobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate (Compound 2); Calculated for
21 C35HsoNloo6 MH+: 707.9, Found: MH~: 707.7;
cis- l ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(4-piperidylmethylcarbamoyl)-1-piperazinecarboxylate (Compound 3); Calculated for
24 C34H53N,006: MHt: 698.9, Found: MH+: 699.7;
cis-1,5-cyclooctylene 4-(trans-4-arninocyclohexylmethylcarbamoyl)-
1-pi~ d~inecarboxylate 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
27 (Compound 4); Calculated for C35H55N,006: MH+: 712.9, Found: MH+: 713.6;
cis- l ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- l -piperazinecarboxylate
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3-piperid-4-ylpropylcarbarnoyl-l-piperazinecarboxylate (Compound 5); Calculated for
C36H58NI0O6: MH+: 727.9 Found: MH+: 727.9;
3 4-[4-(2-piperid-4-ylethylcarbamoyl)piperazin- 1 -ylcarbonyl]benzyl
4-(4-guanidinobenzylcarbamoyl)-l -pi~ ecarboxylate (Compound 6); Calculated for
C34H48N,0O5: MH+: 677.8 Found: MH+: 677.6;
6 4-~4-(3-piperid-4-ylpropylcarbamoyl)piperazin- l -ylcarbonyl]benzyl
4-(4-guanidinobenzylcarbarnoyl)-l-piper~7.inec~rboxylate (Compound 7); Calculated for
C35H50N,0O5: MH+: 691.9 Found: MH+: 691.5;
9 4-[4-(4-piperid-4-ylbutylcarbamoyl)piperazin-1-ylcarbonyl]benzyl
4-(4-guanidinobenzylcarbarnoyl)-1-piperazinecarboxylate (Compound 8); Calculated for
C36H52N,0O5: MH+: 705.9 Found: MH+: 705.9;
12 4-[4-(4-guanidinobenzylcarbamoyl)piperazin- 1 -ylcarbonyl Ibenzyl
4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate (Compound 9); Calculated for
C34H48N,0O5: MH+: 677.8 Found: MH+: 677.7;
4-[4-(4-guanidinobenzylcarbamoyl)piperazin- 1 -ylcarbonyl]benzyl
4-(3-piperid-4-ylpropylcarbamoyl)-1-piperazinecarboxylate (Compound 10); Calculated for
C35E~50N,005: MH+: 6gl.9 Found: MH+: 691.3;
18 4-[4-(2-piperid-4-ylethylcarbamoyl)piperazin-1-ylcarbonylmethyllbenzyl
4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate (Compound 11); Calculated for
C35H50NI0O5: MH+: 691.9 Found: MH+: 692.1;
21 4- [4-(3 -piperid-4-ylpropylcarbamoyl)piperazin- 1 -ylcarbonylmethyl~benzyl
4-(4-guanidinobenzylcarbamoyl)-1-pilJc~ ecarboxylate (Compound 12); Calculated for
C36H52N,0O5: MHt: 705.9 Found: MH+: 705.6;
24 cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbarnoyl)-1-piperazinecarboxylate
4-(4-methylaminomethylbenzylcarbarnoyl)-1-piperazinecarboxylate (Compound 13); Calculated
for C37H54NI0O6: MHt: 735.9 Found: MH+: 735.7;
27 cis-1,5-cyclooctylene 4-(4-guanidinophenylacetyl)-l-piperazinecarboxylate
4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate (Compound 14); Calculated for
C35H55NgO6 MH+: 698.9, Found: MH~: 698.2;
cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- l -piperazinecarboxylate
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4-(3-piperid-4-ylpropylcarbamoyl)-1-piperazinecarboxylate (Compound 15); Calculated for
C36H57NgO6 MH+: 712.9, Found: MH~: 712.3;
3 cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-l-piper~7inec~rboxylate
4-(4-imidazol-1-ylbutylcarbamoyl)-1-piperazinecarboxylate (Compound 16); Calculated for
C35H53N"O6: MH+: 724.9, Found: MH+: 724.5;
6 cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piper~7incc~rboxylate
4-(4-imidazolin-2-ylaminobutylcarbamoyl)-1-pilx;~ ecarboxylate (Compound 17); Calculated
for C35H56N,2O6: MH+: 741.9, Found: MH+: 741.7;
g cis-1,5-cyclooctyiene 4-(trans-4-aminocyclohexylmethylcarbamoyl)-
1-piperazinecarboxylate 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
(Compound 18); Calculated for C35H56N~0O6: MH+: 713.9 Found: MH+: 714.1;
12 cis- 1,5-cyclooctylene 2-(1 -tert-butryryloxymethoxycarbonylpiperid-4-yl)ethylcarbamoyl-
I-piperazinecarboxylate 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
(Compound 19); Calculated for C4~H66NIoOlo MH+: 872.1, Found: MH+: 871.8;
1 5 cis- I ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-[2-(1-methylpiperid-4-yl)ethylcarbamoyl]-1-piperazinecarboxylate (Compound 20); Calculated
for C36H58NI0O6: MH22+12: 364.0, Found: MH22+/2: 364.3;
18 cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- I-piperazinecarboxylate
4-(3-imidazolin-2-ylaminopropylcarbarnoyl)-1-piperazinecarboxylate (Compound 21);
Calculated for C34H54N,2O6: MH~: 727.9, Found: MH+: 728.0;
21 cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-[1-(1-iminoethyl)piperid-4-ylmethylcarbamoyl]-1-piperazinecarboxylate (Compound 22);
Calculated for C36H57NI~O6: MH+: 740.9, Found: MH+: 740.5;
24 cis- 1,5-cyclooctylene 4-(4-guanidinobenzoylarninomethyl)- 1 -piperidinecarboxylate
4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate (Compound 23); Calculated for
C36H57NgO6 MH+: 712.9, Found: MH+: 711.6;
27 cis-1,5-cyclooctylene 4-(4-amidinobenzylcarbarnoyl)-1-piperazinecarboxylate
2-(1 -methoxycarbonylpiperid-4-yl)ethylcarbamoyl- 1 -piperazinecarboxylate (Compound 24);
Calculated for C37H57NgO8 MH+: 756.9, Found: MH+: 756.7; and
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3-{4-[2-(4-{cis-5-[4-(4-amidinobenzylcarbamoyl)piperazin-1 -ylcarbonyloxy]cyclooctyloxy-
carbonyl }piperazin- 1 -ylcarbonylamino)ethyl]piperid- 1 -ylcarbonyl } propionic acid
3 (Compound 25); Calculated for C39H60N909: MHt: 799.0, Found: MH+: 798.6.
Procee~lin~ as in Example 8 and replacing the isocyanate with an activated ester the
following compounds of Formula I were ~lep~ed:
6 cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -pipera_inecarboxylate
4-imidazol-4-ylacetyl~ c~ ecarboxylate (Compound 26); Calculated for
C32H46NI006: MH+: 667.8, Found: MH+: 667.7;
9 cis- 1,5-cyclooctylene 4-(4-guanidinoben_ylcarbamoyl)- 1 -piperazinecarboxylate
4-(E-3-imidazol-4-ylacryloyl)-1-piperazinecarboxylate (Compound 27); Calculated for
C33H46N,006: MH+: 679.9, Found: MH+: 679.8;
12 cis- 1,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(3-imidazol-4-ylpropionyl)-1-piperazinecarboxylate (Compound 2~); Calculated for
C33H48N,006: MH+: 681.8, Found: MH~: 681.7;
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl~- 1 -piperazinecarboxylate
4-(5-imidazol- 1 -ylvaleryl)- I -pipera inecarboxylate (Compound 29~; Calculated for
C35H52N~006: MH+: 709.9, Found: MH+: 709.5;
18 cis- I ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(6-imida_ol-1-ylhexanoyl)-1-piperazinecarboxylate (Compound 30); Calculated for
C36H54N~006: MH+: 723.9, Found: MH+: 723.4;
21 cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbarnoyl)- 1 -piper~7.inec~rboxylate
4-(4-imidazol-1 -ylmethylphenylacetyl)~ ela;~inecarboxylate (Compound 31); Calculated for
C39H52NI006: MH+: 757.9, Found: MH+: 757.2;
24 cis- 1,5-cyclooctylene 4-(4-guanidinoben~ylcarbarnoyl)- 1 -piperazinecarboxylate
4-(4-imidazol-1-ylmethylbenzoyl)-1-~ip~ ec~rboxylate (Compound 32); Calculated for
C38H50N~006: MH+: 743.9, Found: MH+: 743.7;
27 cis- 1,5-cyclooctylene 4-(4-guanidinoben_ylcarbamoyl)- 1 -piper~inçç~rboxylate
4-(3-imi~1~701-1-ylmethylbenzoyl)-l-piperazinecarboxylate (Compound 33); Calculated for
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-41 -
C38H50N,006: MH+: 743.9, Found: MH+: 743.6;
cis-1,5-cyclooctylene 4-(4-gll~ni~linobenzylcarbamoyl)-1-piperazinecarboxylate
3 4-(7-imidazol-1-ylheptanoyl)-1-piperazinecarboxylate (Compound 34); Calculated for
- C37H56N~006: MH+: 737.9, Found: MH+: 737.6;
cis- 1 ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate6 4-[6-(2-methylimidazol-1-yl)hexanoyl]-1-pipera~inecarboxylate (Compound 35); Calculated for
C37Hs6N~0o6 MH+: 737.9, Found: MH+: 737.3;
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
9 4-(4-imidazol-1-ylphenoxyacetyl)-1-piperazinecarboxylate (Compound 36); Calculated for
C38H50N~oO7 MH+: 759.9, Found: MH+: 759.3;
cis- I ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate12 4-[6-(4-methylimidazol- 1 -yl)hexanoyl]- l -piperazinecarboxylate and cis- 1,5 -cyclooctylene
4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-[6-(5-methylimidazol-1-yl)hexanoyl]-1-~ )eld~hlecarboxylate as a mixture (Compound 37);
15 Calculated for C37H56NI006: MH+: 737.9, Found: MH+: 738.2;
cis- I ,5 -cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piper~7inf c~rboxylate
4-(4-piperid-4-ylbutyryl)-1-pipeld inecarboxylate (Compound 38); Calculated for
18 C36H5,N906: MH+: 712.9 Found: MH+: 712.4;
cis- 1,S-cyclooctylene 4-(4-guanidinophenylacetyl)- l -piperazinecarboxylate
4-(4-piperid-4-ylbutyryl)-1-piperazinecarboxylate (Compound 39); Calculated for
21 C36H56N806: MH+: 697.9, Found: MH+: 697.5;
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperzl7inec~rboxylate4-(2-piperid-4-ylethyl)(methyl)carbamoyl-1-~ipe~ ecarboxylate (Compound 40); Calculated
24 for C36H58N,006: MH+: 727.9, Found: MH+: 727.6;
cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -pipera~inecarboxylate
4-(2-piperid-4-ylethyl)(methyl)carbamoyl- 1 -piperazinecarboxylate (Compound 41); Calculated
27 for C36H57Ng06: MH+: 712.9, Found: MH+: 712.7;
- cis- 1,5 -cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(2-piperid-4-ylethoxycarbonyl)-1-piperazinecarboxylate (Compound 42); Calculated for
30 C35H55N907: MH+: 714.9, Found: MH+: 714.5;
_,
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cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
4-(4-imidazol-1-ylphenylacetyl)-1-piper~inecarboxylate (Compound 43); Calculated for
3 C38H50N~0O6: MH+: 743.9, Found: MH+: 743.6;
cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piper~inecarboxylate
4-(6-imidazol-1-ylhexanoyl)-1-piperazinecarboxylate (Compound 44); Calculated for
6 C36H53NgO6 MH+: 708.9, Found: MH+: 708.8;
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(3-pyrid-4-ylthiopropionyl)-1-piper~7inec~rboxylate (Compound 45); Calculated for
9 C35H4gN9O6 MH+: 724.9, Found: MH+: 724.4;
cis-1,5-cyclooctylene 4-~4-guanidinobenzylcarbonyl)-1-piperazinecarboxylate
4-pyrid-4-ylthioacetyl-1-piperazinecarboxylate (Compound 46); Calculated for~2 C34H47N9O6: MH+: 710.9, Found: MH+: 710.8;
cis- I ,5 -cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-(3-pyrid~-ylthiopropionyl)-1-piperazinecarboxylate (Compound 47); Calculated for~5 C35H48N8O6: MH+: 709.g, Found: MH+: 709.3;
cis- I ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(6-imidazol-4-ylhexanoyl)-1-piperazinecarboxylate (Compound 48); Calculated for
8 C36H54N,0O6: MH+: 723.9, Found: MH+: 723.5;
cis- 1,S-cyclooctylene 4-(benzoimidazol-6-ylcarbonyl)- 1 -piperazinecarboxylate
4-(4-guanidinobenzylcarbamoyl)-1-pip~lazinecarboxylate (Compound 49); Calculated for
21 C35H47N,0O6: MH+: 703.8, Found: MH+: 703.4;
cis- I ,5-cyclooctylene 4-(4-amidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(6-imidazol-1-ylhexanoyl)-1-pip~.~i,~ecarboxylate (Compound 50); Calculated for~4 C36H53NgO6 MH+: 708.9, Found: MH+: 708.6;
cis- I ,5-cyclooctylene 4-(4-amidinobenzoylaminomethyl)- 1 -piperidinecarboxylate
4-(6-imi~7O1-4-ylhexanoyl)-1-piperazinecarboxylate (Compound 51); Calculated for~7 C3,H54N8O6: MH+: 707.9, Found: MH+: 707.5;
cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-(6-imidazol-4-ylhexanoyl)-1-piperazinecarboxylate (Compound 52); Calculated for
30 C35H52N,0O6: MH+: 708.9 Found: MH+: 708.4;
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-43-
cis- l ,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-pyrid-4-ylcarbamoylacetyl-l-piper~7in~c~rboxylate (Compound 53); Calculated for
3 C35H4,N907: MH+: 706.8, Found: MH+: 706.3;
- cis- 1,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(3-pyrid-4-ylarninopropionyl)-1-piperazinecarboxylate (Compound 54); Calculated for
6 C3sH50N~0o6 MH+: 707.9, Found: MH+: 707.3;
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
3-[pyrid-4-yl(tert-butoxycarbonyl)arnino]propionyl-1-piperazinecarboxylate (Compound 55);
9 Calculated for C4~Hs8N,0O8: MH22+/2: 404.5, Found: MH22+/2: 404.2;
cis- l ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(3-piperazin-1-ylcarbonylpropionyl)-1-piperazinecarboxylate (Compound 56); Calculated for
2 C35Hs4N,007: MH+: 727.9, Found: MH+: 727.5;
cis- l ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-piperid-1-ylcarbonylaminoacetyl-1-piperazinecarboxylate (Compound 57); Calculated for
5 C35H54NloO,: MH+: 727.9, Found: MH+: 727.5;
cis- 1 ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate4-(5-imidazol~-ylvaleryl)-1-piperazinecarboxylate (Compound 58); Calculated for
18 C35H52N~0O6: MH+: 708.9, Found: MH+: 709.4;
cis- l ,5-cyclooctylene 4-(4-amidinobenzoylaminomethyl)- 1 -piperidinecarboxylate
4-(3-piperazin-1-ylcarbonylpropionyl)-1-piperazinecarboxylate (Compound 59); Calculated for
21 C36H54N8O7: MH+: 711.9, Found: MH+: 711.4;
cis- 1,S -cyclooctylene 4-(4-atnidinobenzoylaminomethyl)- 1 -piperidinecarboxylate
4-piperid-4-ylcarbonylaminoacetyl-1-piperazinecarboxylate (Compound 60); Calculated for
24 C36H54N8O7: MH+: 711.9, Found: MH+: 711.4;
cis- 1,5-cyclooctylene 4-~3-(2-aminopyrimidin-5-yl)propionyl]- 1 -piper~in~c~rboxylate
4-(4-guanidinobenzylcarba~noyl)-1-piperazinecarboxylate (Compound 61); Calculated for
27 C34H49N"O6: MH+: 708.8, Found: MH+: 708.4;
cis- 1,5-cyclooctylene 4-[3-(6-aminopyrid-3-yl)propionyl]- 1 -piperazinecarboxylate
4-(4-guanidinobenzylcarbamoyl)-1-~i~e~Ginecarboxylate (Compound 62); Calculated for
30 C35H50N'0O6: MH+: 707.8, Found: MH+: 707.4;
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cis- I ,5 -cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-[4-(4-pyrid-4-ylthio)butyryl]-l-piperz~7inec~rboxylate (Compound 63); Calculated for
3 C36H5lN9O6: MH+: 738.9, Found: MH+: 738.4;
cis- 1,5-cyclooctylene
4-[3-(2-amino-2,4-dioxo- 1,2,3,4-tetrahydropyrimidin-5-yl)propionyl]-1 -piperazinecarboxylate
6 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate (Compound 64); Calculated for
C34H48N,0O8: MH+: 725.8, Found: MH+: 725.2;
cis- 1,5 -cyclooctylene 4-(4-amidinobenzylcarbamoyl)- 1 -pipel ;~inecarboxylate
9 4-(4-piperid-4-ylbutyryl)-l-piperazinecarboxylate (Compound 65); Calculated for
C36H56N8O6: MH+: 697.9, Found: MH+: 697.4;
cis- 1,5-cyclooctylene 4-(4-amidinobenzoylaminomethyl)- 1 -piperidinecarboxylate2 4-(4-piperid-4-ylbutyryl)- 1-piperazinecarboxylate (Compound 66); Calculated for
C37H57N706: MH+: 696.9, Found: MH+: 696.4;
cis- 1,5-cyclooctylene 4-(1 -arnidinopiperid-4-ylacetyl)- 1 -piperidinecarboxylate
5 4-(6-imidazol-l-ylhexanoyl)-1-piper~inec~rboxylate (Compound 67); Calculated for
C35H57N9O6: MH+: 700.9, Found: MH+: 700.5;
cis- 1,5-cyclooctylene 4-(1 -arl1idino-4-piperidylacetyl)- 1 -piperazinecarboxylate
8 4-(4-piperid-4-ylbutyryl)-1 -piperazinecarboxylate (Compound 68); Calculated for
C35H60N8O6: MH+: 689.9, Found: MH+: 689.4;
cis- 1,5-cyclooctylene 4-(1 -amidino-4-piperidylacetyl)- 1 -piperazinecarboxylate
21 4-(6-imidazol-1-ylhexanoyl)-1-pip~ ecarboxylate (Compound 69); Calculated for
C35H57NgO6 MH+: 700.9, Found: MH+: 700.4;
cis- 1,5 -cyclooctylene 4-(4-amidinobenzylcarbarnoyl)- 1 -~ipe~ ecarboxylate
24 4-(6-imidazol-4-ylhexanoyl)-1-piperazinecarboxylate (Compound 70); Calculated for
C36H53NgO6 MHt: 708.9, Found: MH+: 708.4;
cis- 1,5 -cyclooctylene 4-(4-amidinobenzoylaminomethyl)- 1 -piperidinecarboxylate
27 4-(6-imidazol-4-ylh~noyl)-1-piperazinecarboxylate (Compound 71); Calculated for
C37H54N8O6: MH+: 707.9, Found: MH+: 707.4;
cis- 1,5 -cyclooctylene 4-(4-arnidinophenylacetyl)- 1 -pipel d~inecarboxylate
4-(6-imidazol-1-ylhexanoyl)-l-pi~ inecarboxylate (Compound 72); Calculated for
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C36H52N8O6: MH+: 693.9, Found: MH+: 693.4;
cis- 1,5-cyclooctylene 4-(4-amidinophenylacetyl)- 1 -piper~7inec~rboxylate
3 4-(4-piperid-4-ylbutyryl)-1-piperazinecarboxylate (Compound 73); Calculated for
- C36H5~N,O6: MH+: 682.9, Found: MH+: 682.4;
cis- 1,5-cyclooctylene 4-(4-amidinophenylacetyl)- 1 -piperazinecarboxylate
6 4-(6-imidazol-4-ylhexanoyl)-1-pipeld~ ecarboxylate (Compound 74); Calculated for
C36H52N8O6: MH+: 693.9 MH+: 693.4; and
cis- 1,5-cyclooctylene 4-(4-amidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
9 4-[4-(2-(1 -tert-butylcarbonyloxymethoxycarbonyl)piperid-4-ylethylcarbamoyl)-
l-piperazinecarboxylate (Compound 7S); Calculated for C42H65N9O~o MH+: 857.0
Found: MH+: 856.6.
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EXAMPLE 9
cis- I ,5-cyclooctylene di(4-tert-butoxycarbonyl- 1 -,~)ipeldzillecarboxylate)
3 cis-1,5-Cyclooctylene di(chloroformate) (3.69 g, 13.7 rnmol.), prepared as in Example 5,
and DIEA (7.2 mL, 41 mmol.) were taken into DMF (25 mL) and tert-butyl
l-piperazinecarboxylate (5.1 g, 27.4 mmol.) was added. The mixture was stirred 12 hours at
6 room temperature and then concentrated in vacuo giving a semi-solid residue. The residue was
partitioned between dichloromethane (50 mL) and water (50 mL) and the dichloromethane layer
was washed with 0.1N aqueous hydrochloric acid (2x, 25 mL), dried (MgS04) and filtered.
9 concell~ldling in vacuo gave cis-l,S-cyclooctylene di(4-tert-butoxycarbonyl-1-piperazinecarboxylate) as an amorphous solid; IH-NMR (300MHz, CDC13): 4.80 (m, 2H), 3.40
(br s, 16H), 2.00-1.40 (m, 12H), 1.40 (s, 18H).
12 EXAMPLE 10
cis- 1 ,5-cyclooctylene di[4-(2-piperid-4-ylethylcarbamoyl)- 1 -piperazinecarboxylate]
(Compound 76)
The following is the preparation of a compound of Formula I in which R' and R2 each are
2-piperid-4-ylethyl, Xl and X9 each are -NHC(O)-, x2 and x8 each are 1,4-piperazinylene, X3 and
X7 each are -C(O)O-, X4 and x6 each are a covalent bond and X5is cis-1,5-cyclooctylene.
~8 cis-l,S-Cyclooctylene di(4-ter~-butoxycarbonyl-1-piperazinecarboxylate) (47.9 mg,
0.088 mmol.), prepared as Example 9, was treated with T~A (I mL) neat for 10 minutes giving a
colorless oil. The mixture was concentrated in vacuo and the residue was triturated with ethyl
21 ether (2x, 5 mL) and repeatedly dried in vacuo giving an amorphous solid. The solid residue was
taken into DMF (5 mL) and DIEA (100 mL, 0.5 mrnol.) and then tert-butyl
4-(2-isocyanatoethyl)-1-piperidinecarboxylate (460 mL, 0.39 M in DMF, 0.18 mmol.) was added
24 to the solution. The mixture was stirred 12 hours and concentrated in vacuo. The residue was
triturated with water (2x, 5 mL) and dried in vacuo giving a yellow solid. The solid was treated
with TFA (2 mL) and the mixture was concentrated in vacuo. The residue was taken into water.
27 Purifying from the aqueous mixture by p le~)ar~live reverse phase HPLC followed by
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lyophilli7~tion gave cis-1,5-cyclooctylene di[4-(2-piperid-4-ylethylcarbamoyl)-
1-piperazinecarboxylate] as a colorless amorphous solid; Electrospray LRMS: Calculated for
3 C34H6~N8O6: MH+: 677.9, Found: MHt: 677.6.
i
Proceeding as in Example 10 and sub~liluLi~lg different starting materials
cis- l ,5-cyclooctylene di[4-(4-methylaminomethylbenzylcarbamoyl)- 1 -piperazinecarboxylate]
6 (Compound 77) was plel~aled; Calculated for C38H56N8O6: MH+: 721.9, Found: MH+: 721.7.
Proceeding as in Example 10 and replacing the isocyanate with an activated ester the
following compounds of Forrnula I were prepared:
9 cis- 1,5-cyclooctylene di [4-(4-piperid-4-ylbutyryl)- 1 -piperazinecarboxylate]
(Compound 78); Calculated for C36H62N6O6: MH+: 675.9, Found: MH+: 675.6; and
para-dimethylenephenylene di[4-(4-piperid-4-ylbutyryl)-1-piperazinecarboxylate]
2 (Compound 79); Calculated for C36H56N6O6: MH+: 669.9, Found: MH+: 669.4.
EXAMPLE 11
ter~-Butyl 4-(3 -imidazol - I -ylpropylcarbamoyl)- I -piperazinecarboxylate
tert-Butyl 4-chlorocarbonyl-1 -piper~inec~rboxylate (188 mg, 0.76 mmol.), prepared as
in Example 3, was taken into dichloromethane (10 mL) and DIEA (150 mL, 0.86 mmol) was
added. 1-(3-Aminopropyl)imidazole (100 mL, 0.84 mmol) was added by syringe and the
8 mixture was stirred 12 hours. Dichloromethane (10 mL) was added to the mixture and the
organic layer was washed with water ( I x, 10 mL), dried (MgSO4) and filtered. Concentrating
gave tert-butyl 4-(3-imidazol-1-ylpropylcarbamoyl)-1-piperazinecarboxylate (230 mg,
21 0.68 rnmol, 90% yield) as a colorless oil; 'H-NMR (300MHz, DMSO-d6): 7.60 (s, IH), 7.20 (s,
IH), 6.85 (s, IH), 6.60 (tr, IH), 3.95 (tr, 2H), 3.30 (s, 8H), 3.00 (q, 2H), 1.80 (m, 2H), 1.40 (s,
9H).
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EXAMPLE 12
tert-Butyl 4-aminomethyl- I-benzenecarbamate hydrochloride
3 4-Aminobenzylamine ~5.56 g, 45.6 mmol.) was taken into water (45 mL) and citric acid
(9.63 g, 50 mmol) was added to the solution. Di-tert-butyl dicarbonate (9.94 g, 45.5 mmol) in
dioxane (20 mL) was added dropwise to the solution and the mixture was stirred 48 hours at
6 room temperature giving a yellow suspension. The suspension was filtered and the aqueous
solution was basifled with excess solid sodium carbonate and extracted with ethyl acetate (3x,
35 mL). The combined extracts were washed with saturated aqueous sodium chloride, dried
9 (MgSO4), f1ltered and concentrated in vacuo giving a white solid. The solid was taken into
methanol (30 mL), the solution was acidified with hydrogen chloride in dioxane (4M, 8.4 mL,
33.6 mmol.) and then ethyl ether (100 mL) was added giving a suspension. The particulate
12 matter was isolated by filtration. Drying in vacuo gave tert-butyl
4-aminomethyl-l-ben7~nec~rbamate hydrochloride (7.2 g, 27.8 mmol, 61% yield) as a colorless
solid; 'H-NMR (300MHz, DMSO-d6): 9.43 (s, lH), 8.20 (br s, 3H), 7.40 (dd AB, 4H), 3.92 (m,
2H), 1.50 (s, 9H).
EXAMPLE 13
tert-Butyl 4-isocyanatomethyl-1-benzenecarbamate:
18 tert-Butyl 4-aminomethyl-l-ben7.Pnec~rbamate hydrochloride (3.39 g, 13.1 mmol),
prepared as in Example 12, was taken into dichloromethane (120 mL) at 0~C and pyridine
(4.3 mL, 53 mmol) and triphosgene (1.3 g, 4.4 mmol) was added. The mixture was allowed to
21 warm to room temperature over 30 minutes and aqueous hydrochloric acid (0.5N, 100 mL) was
added. The organic layer was dried (MgSO4) and filtered. Conce,ll,d~ g gave tert-butyl
4-isocyanatomethyl-1-benzenecarbamate (2.7 g, 11 mmol, 84% yield) as a yellow solid;
24 'H-NMR (300MHz, CDCI3): 7.29 (dd AB, 4H), 6.55 (br s, lH), 4.40 (s, 2H), 1.55 (s, 9H).
, . . . .
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EXAMPLE 14
cis-1,5-Cyclooctylene 4-(4-arninobenzylcarbamoyl)-1-piperazinecarboxylate
3 4-(3 -imidazol- 1 -ylpropylcarbamoyl)- l -piper~i n~c~rboxylate
(Compound 80)
The following is the preparation of a compound of Formula I in which R' is
6 4-aminobenzyl, R2 is 3-imidazol-l-ylpropyl, Xl and X9 each are -NHC(O)-, x2 and X~ each are
1,4-piperazinylene, X3 and X7 each are -C(O)O-, X4 and x6 each are a covalent bond and X5 iS
cis- 1 ,5-cyclooctylene.
g (a) tert-Butyl 4-(3-imidazol-1-ylpropylcarbamoyl)-1-piperazinecarboxylate (225 mg,
0.67 mmol), prepared as in Example 1 1, was treated with TFA (1 mL) neat for 10 minutes. The
mixture was concentrated in vacuo and the residue was taken into dichloromethane (10 mL) and
12 an excess of DIEA (1.0 mL) was added to the solution. cis-1,5-Cyclooctylene chloroformate
4-~er~-butoxycarbonyl-1-piperazinecarboxylate (279 mg, 0.67 mmol), prepared as in Example 6,
in dichloromethane (5 mL) was added to the solution and the mixture was stirred for 1 hour.
1 5 ~dditional dichloromethane ( 10 mL) was added and the organic layer was washed with saturated
aqueous sodium bicarbonate (lx, 10 mL), dried (MgSO4) and filtered. Concentrating gave crude
cis- 1 ,5-cyclooctylene 4-(3-imidazol- 1 -ylpropylcarbamoyl)- 1 -piperazinecarboxylate
18 tert-butoxycarbonyl- 1 -piperazinecarboxylate adduct as a colorless foam.
(b) The adduct prepared in Part (a) was treated with TFA (l mL) neat for 10 minutes and
then the mixture was concentrated in vacuo. The residue was taken into DMF (10 mL) and an
21 excess of DIEA (1.5 mL) and tert-butyl 4-isocyanatomethyl-1-benzenecarbamate (165 mg,
0.67 mmol.), prepared as in Example 13 were added. The mixture was stirred 12 hours and
concentrated in vacuo. The residue was treated with TFA neat and mixture was concentrated
24 in vacuo. The residue was taken into water (15 mL) and the aqueous solution was extracted with
ethyl ether (lx, 15 mL). The aqueous layer was basified with l.OM aqueous sodium hydroxide
and then extracted with dichloromethane. The dichloromethane was dried (MgSO4) and filtered.
27 Concentrating in vacuo gave cis-1,5-cyclooctylene 4-(4-aminobenzylcarbamoyl)-
l-piperazinecarboxylate 4-(3-imidazol-1-ylpropylcarbamoyl)-1-piperazinecarboxylate as a
colorless foam; ~H-NMR (300MHz, CDCI3): 7.45 (s, lH), 7.10 (d, 2H), 7.05 (s, lH), 6.90 (s,
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IH), 6.60 (d, 2H), 4.85-4.70 (m, 4H), 4.30 (d, 2H), 4.00 (tr, 2H), 3.50-3.30 (m, 18H), 2.00 (m,
2H), 1.90-1.50 (m, 12H).
3 Proceeding as in Example 14 and substituting different starting materials the following
compounds of Formula I were prepared:
cis- I ,S-cyclooctylene 4-(4-aminobenzylcarbamoyl)- 1 -piperazinecarboxylate
6 4-(2-pyrid-4-ylethylc~rbamoyl)- 1 -piper~7in~c~rboxylate (Compound 81 );
cis- 1 ,S-cyclooctylene 4-(4-aminobenzylcarbarnoyl)- 1 -piperazinecarboxylate
4-(3-piperid-4-ylpropyl)-1-piperidinecarboxylate (Compound 82); and
9 cis- 1 ,S-cyclooctylene 4-(4-aminobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(~-piperid-4-ylbutyl)- 1 -piperidinecarboxylate (Compound 83 ).
EXAMPLE 15
1 2 cis- 1 ,S-Cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(3-imill~7. )1-l-ylpropylcarbarnoyl)-l-piperazinecarboxylate
(Compound 84)
The following is the plel,al~ion of a compound of Formula I in which R~ is
4-guanidinobenzyl, R2 is 3-imidazol-1-ylpropyl, X' and X9 each are -NHC(O)-, x2 and x8 each
are 1,4-piperazinylene, X3 and X7 each are -C(O)O-, X4 and x6 each are a covalent bond and X5
1 8 is cis- 1 ,S-cyclooctylene.
cis- 1 ,5-Cyclooctylene 4-(4-aminobenzylcarbamoyl)- 1 -pip~ inecarboxylate
4-(3-imidazol-1-ylpropylcarbamoyl)-1-piperazinecarboxylate, prepared in Exarnple 14, was
21 taken into methanol and ethyl ether and an excess of hydrogen chloride (4M in dioxane) was
added. The mixture was concentrated and dried in vacuo. An excess of cyanamide (1.0~) was
added and the mixture was heated at 65 ~C for two hours giving a yellow solution. The mixture
24 was allowed to cool to room temperature and triturated with ethyl ether (3x, 10 mL). The
insoluble residue was taken into water. Purifying from the aqueous mixture by preparative
reverse phase HPLC gave cis- 1 ,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-
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1-pipel~in~c~rboxylate 4-(3-imidazol-1-ylpropylcarbamoyl)-1-piperazinecarboxylate as a
colorless amorphous solid; Electrospray LRMS: C~lculated for C34Hs,Nl,O6: MH+: 710.9,
3 Found: MH+: 710.6.
Proceeding as in Example 15 and substituting different starting materials the following
compounds of ~ormula I were prepared:
6 cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- l -piperazinecarboxylate
4-(2-pyrid-4-ylethylcarbamoyl)-1-pil~ldzinecarboxylate (Compound 85); Calculated for
C35H50NloO6 MH+: 707.9 Found: MH+: 707.6;
9 cis- 1,5 -cyclooctylene 3 -piperid-4-ylpropyl- 1 -piperidinecarboxylate4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate (Compound 86); and
cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
12 4-piperid-4-ylbutyl-1-piperidinecarboxylate (Compound 87); Calculated for
C37H60N8O5: MH~: 697.9, Found: MH+: 697.7.
EXAMPI,E 16
cis-1,5-cyclooctylene chloroformate 4-benzyloxycarbonyl-1-piperazinecarboxylate
Benzyl 1-pipera7.inec~rboxylate (1.0 g, 4.53 mmol, 1.0 equiv) and DIEA (0.88 mL,4.9g mmol, 1.1 equiv) in dichloromethane (25 mL) was added dropwise to cis- 1 ,5-cyclooctylene
18 di(chloroformate) (1.2 g, 4.53 mmol, 1.0 equiv), prepared as in Example 5, in dichloromethane
(25 mL) at 0~C. The reaction mixture was stirred for 22 hours while allowing to warm to room
temperature. The mixture was partitioned between dichloromethane, 0.05N aqueous hydrochloric
21 acid and saturated aqueous sodium chloride. The organic layer was dried (Na2SO4) over sodium
sulfate and concentrated. Purifying the residue by flash column chromatography, eluting with 20
and 30% ethyl acetate in hexanes, gave cis-1,5-cyclooctylene chloroformate
24 4-benzyloxycarbonyl-1-piperazinecarboxylate (0.81 g, 1.81 mmol, 40%) as a yellow oil;
IR: 2939 (s), 2863 (m), 1770 (s),1732 (s),1696 (s); 'H NMR (300 MHz, CDCI3): 7.35 (s, 5 H),
5.15 (s, 2 H), 4.95 (m,1 H),4.75 (m, 1 H),3.45 (s,8 H), 1.50 - 2.05 (m, 12 H).
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EXAMPLE 17
I -~cis-5-(4-Benzyloxycarbonylpip~ - 1 -ylcarbonyloxy)cyclooctyloxycarbonyl]-
3 4-piperidinecarboxylic acid
Isonipecotic acid (75 mg, 0.58 mmol, 1.1 equiv) and DIEA (0.23 mL, 1.33 mmol, 2.5
equiv) were added to cis- 1,5-cyclooctylene chloroformate 4-benzyloxycarbonyl-
6 1-pipe~ 1ecarboxylate (0.24 g, 0.53 mmol, 1.0 equiv), prepared as in Example 16, in
dichloromethane (lO mL) at 0~C giving a white suspension. The suspension was stirred for
18 hours while allowing to warm to room temperature. The reaction mixture was partitioned
9 between dichloromethane and 0.05N aqueous hydrochloric acid. Concentrating the organic layer
gave crude l-[cis-5-(4-benzyloxycarbonylpiperazin-1-ylcarbonyloxy)cyclooctyloxycarbonyl]-
4-piperidinecarboxylic acid (0.36 g) as a colorless oil; 'H NMR (300 MHz, CDCl3): 7.35 (s, 5
12 H),5.15(s,2H),4.75(m,2H),3.90(m, lH),3.45(s,8H)~2.75(m, lH),2.50(m,3H),
1.50-1.90 (m, 16 H).
EXAMPLE 18
1 5 cis- 1,5-Cyclooctylene 4-benzyloxycarbonyl- 1 -piperazinecarboxylate
4-[2-(1 -terf-butoxycarbonylpiperid-4-yl)ethylcarbamoyl]- 1 -piperidinecarboxylate
1 -Hydroxybenzotriazole hydrate (80 mg, 58.3 mmol, 1.1 equiv), tert-butyl
8 4-(2-aminoethyl)- 1 -piperidinecarboxylate hydrochloride (0.14 g, 0.53 mmol, 1.0 equiv) and
4-methylmorpholine (0.15 mL, 1.33 mmol, 2.5 equiv) were added to a solution of crude
1 -[cis-5-(4-benzyloxycarbonylpiperazin- 1 -ylcarbonyloxy)cyclooctyloxycarbonyl]-
21 4-piperidinecarboxylic acid (0.36 g, 0.53 mol, 1.0 equiv), prepared as in Example 17, in DMF
(5 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.13 g, 0.66 mmol,
1.25 equiv) was added to the reaction mixture at 0 ~C. The solution was stirred for 1.5 hours at
24 0 ~C and for 3 days at 23 ~C. The reaction mixture was partitioned between dichloromethane,
0.05N aqueous hydrochloric acid, saturated aqueous sodium bicarbonate, water (two portions)
and saturated aqueous sodium chloride. The organic layer was dried (Na2SO4) and concentrated.
27 Purifying from the residue by flash column chromatography, eluting with 3% methanol in
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dichloromethane, gave cis-1,5-cyclooctylene 4-benzyloxycarbonyl-1-piperazinecarboxylate
4-[2-(1 -tert-butoxycarbonylpiperid-4-yl)ethylcarbamoyl]- 1 -piperidinecarboxylate (0.15 g,
3 0.2 mmol, 37% over two steps) as a yellow oil; 'H NMR (300 MHz, CDCl3): 7.35 (s, 5 H), 5.15
(s,2H),4.80(m,2H),4.10(m,4H),3.45(m, lOH),3.30(m,2H),2.70(m,2H), 1.50-l.90(m,
23 H), 1.45 (s, 9 H); Electrospray LRMS: calcd for C40H62NsOg (MH+): 756.97, obtained: 757Ø
6 EXAMPLE 19
cis-1,5-Cyclooctylene 1-piperazinecarboxylate
4-[2-(1 -tert-butoxycarbonylpiperid-4-yl)ethylcarbamoyl]- 1 -piperidinecarboxylate
g Ethanol (3 mL) was added to cis-1,5-cyclooctylene 4-benzyloxycarbonyl-
I-piper~7.inec~rboxylate 4-[2-(1-tert-butoxycarbonylpiperid-4-yl)ethylcarbamoyl]-
1-piperidinecarboxylate (0.15 g, 0.20 mmol, l .0 equiv), prepared as in Example 18, and
2 5% palladium on carbon (75 mg, 0.50 wt equiv) under nitrogen. The mixture was stirred under
hydrogen (1 atm) for 17 hours at 23 ~C. The reaction mixture was placed under nitrogen and
filtered. Concentrating the filtrate gave cis-1,5-cyclooctylene 1-piperazinecarboxylate
4-[2-(1 -tert-butoxycarbonylpiperid-4-yl)ethylcarbamoyl] - I -piperidinecarboxylate (110 mg,
0.18 mmol, 90%) as a colorless oil; ~H NMR (300 MHz, CDCl3): 5.5 (m, 1 H), 4.9 (m, 2 H), 4.75
(m, 1 H), 4.1 (m, 4 H), 3.45 (m, 4 H), 3.25 (m, 2 H), 2.60-2.85 (m, 8 H), 2.10 (m, 1 H), 1.50-1.95
1 8 (m, 23 H), 1.45 (s, 9 H); Electrospray LRMS: calcd for C32H56N5O7 (MH+): 622.83,
obtained: 622.7.
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EXAMPLE 20
cis-1,5-Cyclooctylene 4-(4-guanidinoben_ylcarbamoyl)-1-piperazinecarboxylate
3 4-(2-piperid-4-ylethylcarbamoyl)- 1 -piperidinecarboxylate
(Compound 88)
The following is the pl~aldlion of a compound of Formula I in which R' is
6 4-guanidinoben~yl, R2 is 2-piperid-4-ylethyl, X' and X9 each are -NHC(O)-, x2 is
1,4-pi~ dzil,ylene, x8 is 4,1-piperidylene, X3 and X7 each are -C(O)O-, X4 and x6 each are a
covalent bond and X5iS cis-1,5-cyclooctylene.
9 Triphosgene (30 mg, 0.10 mrnol, 0.58 equiv) and pyridine (30 mL, 0.39 mmol, 2.1 equiv)
were added to cis-1,5-cyclooctylene 1-piper~7in~-c~rboxylate
4-~2-(1-tert-butoxycarbonylpiperid-4-yl)ethylcarbarnoyl ~ piperidinecarboxylate (0.1 1 g,
12 O.lg rnmol, l.0 equiv), prepared as in Example 19, in dichloromethane (2 mL) at 0~C. The
reaction mixture was stirred 3 hours at 0~C. The mixture was partitioned betweendichloromethane, 0.05N aqueous hydrochloric acid and saturated aqueous sodium chloride. The
15 organic layer was dried (Na~SO4) and concentrated giving a brown oil residue.4-Guanidinobenzylamine dihydrochloride (43 mg, 0.20 mmol, l . l equiv) and DIEA (0.16 mL,
0.90 rnmol, 5.0 equiv) in DMF (2 mL) were added to the residue giving a suspension. The
8 suspension was stirred 18.5 hours at 23 ~C and concentrated. The residue was taken into 50%
TFA in dichloromethane (4 mL) and the mixture was stirred 45 minutes at 23 ~C. The reaction
mixture was concentrated and the residue was triturated with ether and dried in vacuo. Purifying
21 from the residue by prel)~d~ e reverse phase HPLC and Iyophilli7:~tion gave
cis- 1 ,S-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- l -piperazinecarboxylate4-(2-piperid-4-ylethylcarbamoyl)-1-piperidinecarboxylate as a colorless amorphous solid;
24 Electrospray LRMS: calcd for C36H58N9O6 (MH+): 712.91, obtained: 712.g
Procee~ling as in Exarnple 20 and substituting different starting materials the following
compounds of Formula I were p~ ,aled:
27 cis- I ,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piper~7.inec~rboxylate
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4-(3-imidazol-4-ylpropylcarbarnoyl)-1-piperazinecarboxylate (Compound 89); Calculated for
C34HsoNloo6 MH+: 695.9, Found: MH+: 695.4;
3 cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piper~7inec~rboxylate
4-(2-imidazol-4-ylethylcarbarnoyl)-1-piperazinecarboxylate (Compound 90); Calculated for
C33H48N,~O6: MH~: 681.3, Found: MH+: 680.9;
6 cis- 1,5-cyclooctylene 4-(4-g-l~nirlinophenylacetyl)- 1 -piper~inec~rboxylate
4-(3-imidazol- 1 -ylpropylcarbamoyl)- 1 -pi~uC~ hlecarboxylate (Compound 91); Calculated for
C341~50NIoO6: MH+: 695.9, Found: MH+: 694.9;
9 cis- 1,5-cyclooctylene 4-(4-guanidinophenylacetyl)- 1 -piperazinecarboxylate
4-(4-imidazol-1-ylbutylcarbamoyl)-1-pi~eld~ ecarboxylate (Compound 92); Calculated for
C35Hs3N,0O6: MH+: 709.9, Found: MH+: 709.4; and
12 cis- 1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate
4-(3-imidazol-1-ylpropylcarbamoyl)-1-pipeld;Ginecarboxylate (Compound 93); Calculated for
C35H51N~0O6: MH+: 709.9, Found: MH+: 710.4.
EXAMPLE 21
3-Piperid-4-ylpropionic acid hydrochloride
4-Pyridineacrylic acid hydrochloride (12.0 g; 64.6 mmol) and 1.37 g o~platinum oxide
18 was suspended in acetic acid (80 mL) and hydrogenated 12 hours at 50 to 60 psi. The mixture
was diluted with water and filtered through a pad of celite. The solids were washed with water
~200 mL) and the combined filtrate and washings were concentrated in vacuo giving a white
21 solid. The solid was suspended in a small arnount of methanol and the mixture was diluted with
diethyl ether (200 mL). The particulate matter was isolated by filtration and washed with diethyl
ether and hexane. Air-drying gave 3-piperid-4-ylpropionic acid hydrochloride (11.3 g, 58.1
24 mmol, 90%) as a colorless solid; 'H-NMR (300 MHz; DMSO-d6): 8.75 (br s, 2H), 3.15 (d, 2H),
2.75 (t, 2H) 2.2 (t, 2H), 1.75 (d, 2H), 1.45 (t, 2H), 1.25 (br q, 2H).
EXAMPLE 22
27 3-(1-tert-Butyoxycarbonylpiperid-4-yl)propionic acid
.
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3-Piperid-4-ylpropionic acid (5.07 g; 26.2 mmol), prepared as in Example 21, wasdissolved in 2~ aqueous NaOH (40 mL; 80 mmol). THF (40 mL) and then
3 di(tert-butyl)dicarbonate (6.21 g; 28.4 mrnol) was added giving a suspension. The suspension
was stirred 22 hours, diluted with water and concentrated in vacuo. The residue was washed
with diethyl ether (2x, 100 mL) and the a~ueous phase was acidified to pH 2-3 with l.ON
6 a~ueous KHSO4 and extracted with ethyl acetate (3x, 200 mL). The combined orgarlic phases
were washed with brine and dried (Na2SO4). Concentrating in vacuo gave
3-[4-(1-tert-butoxycarbonyl)-4-piperidyl]propionic acid (6.21 g; 24.1 rnmol, 92%) as a colorless
g oil that crystallized on st~nding; ~H-NMR (300 MHz, CDCl3): 4.10 (br d, 2H), 2.65 (br t, 2H),
2.35 (t, 2H), 1.70-1.50 (m, 3H), 1.45 (s, 9H), 1.20-O.g5 (m, 2H).
EXAMPLE 23
12 tert-Butyl 4-benzyloxycarbonyl-1-piperazinecarboxylate
tert-Butyl 1-piperazinecarboxylate(2.01 g; 10.8mmol)andDIEA(2.0mL; 1.48g; 11.5
mmol) in 50 mL of ice cQld dichloromethane was treated with benzyl chloroformate (2.0 mL;
16 2.39 g; 14.0 mmol). The mixture was stirred for 42 hours and then partitioned between ethyl
acetate and O.SN KHSO4. The aqueous phase was extracted with ethyl acetate and the combined
organic layers were washed with water and brine, dried (MgSO4) and concentrated. Purifying
18 firom the residue by chromatography over silica gel (ethyl acetate:hexane, 1:3) gave tert-butyl
4-benzyloxycarbonyl-1-piperazinecarboxylate (3.33 g; 10.4 mmol, 96%) as acolorless solid;
~H-NMR (300 MHz, CDCl3): 7.35 (br s, 5H), 5.13 (s, 2H), 3.55-3.25 (m, ~H), 1.45 (s, 9H).
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EXAMPLE 24
Benzyl l-piperazinecarboxylate hydrochloride
3 tert-Butyl 4-benzyloxycarbonyl-1-piperazinecarboxylate (1.Olg; 3.16 mmol), prepared as
in Example 23, was suspended in 4 mL of ethyl acetate. The suspension was cooled in an ice
water bath and 4N hydrogen chloride (12 mL in 1,4-dioxane) was added giving a solution. The
6 solution was stirred for 30 minutes on the ice bath and then for 30 minutes at room temperature.
The reaction mixture was diluted with diethyl ether (75 mL) giving a precipitate. The precipitate
was isolated by filtration and washed with diethyl ether. Drying in vacuo gave benzyl
9 l-piperazinecarboxylate hydrochloride (740 mg; 2.78 mmol, 88%) as a colorless solid; ~H-NMR
(300 MHz, DMSO-d6): 9.25 (br s, 2H), 7.33 (s, 5H), 5.06 (s, 2H), 3.58 (br s, 4H), 3.04 (t, 4H).
EXAMPLE 25
12 tert-Butyl 4-[2-(4-benzyloxycarbonylpiperazin-1-ylcarbonylamino)ethyl]-
l -piperidinecarboxylate
3-[4-(1-tert-Butoxycarbonyl)-4-piperidyl]propionic acid (2.16 g; 8.4 mmol), prepared as
in Example 227 in dry benzene (28 mL) was treated with triethylamine (1.35 mL; 951 mg; 9.40
mrnol) and diphenylphosphoryl azide (2.05 mL, 2.62 g; 9.53 mmol). The reaction mixture was
gradually heated to reflux and kept at reflux for 3.5 hours. The mixture was cooled to room
18 tell,p~ldlule then added dropwise to a suspension of benzyl l-piperazinecarboxylate
hydrochloride (2.44 g; 9.19 mmol), prepared as in Example 24, and triethylamine (1.40 mL;
1.02g; 10.0 mmol) in dry dichloromethane (10 mL). The reaction mixture was stirred for 43
21 hours and diluted ~,vith ethyl acetate and 0.5N KHSO4. The organic layer was washed with water,
aqueous sodium bicarbonate and brine, dried (Na2SO4) and concentrated. Purifying from the
residue by chromatography on silica gel ~ethyl acetate-hexane, 4:1; then pure ethyl acetate) gave
24 ter~-butyl 4-[2-(4-benzyloxycarbonylpiperazin- 1 -ylcarbonylamino)ethyl]- I -piperidinecarboxylate
(3.84 g, 8.1 mmol, 96%) as a white solid; ~H-NMR (300 MHz, CDCI3): 7.35 (s, 5H), 5.15 (s,
2H), 4.50 (br t, I H), 4.05 (br s, 2H), 3.55-3.45 (m, 4H), 3.40-3.30 (m, 4H), 3.25 (q, 2H), 2.65 (t,
27 2H), 1.70 (s, 2H), 1.45 (s, 1 lH), 1.20- 1.00 (m, 2H).
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EXAMPLE 26
cis-1,5-Cyclooctylene chloroformate 4-[2-(4-tert-butoxycarbonylpiperidin-4-yl)ethylcarbarnoyl]-
3I-piper~in~c~rboxylate
/ert-Butyl 4-[2-(4-benzyloxycarbonylpi~el~zill-1-ylcarbonylarnino)ethyl]-
1-piperidinecarboxylate (2.03 g; 4.28 mmol), prepared as in Example 25, and 10%
6 palladium-on-carbon (570 mg) suspended in ethanol (19 mL) was hydrogenated at atmospheric
pressure overnight. The reaction mixture was filtered and the catalyst was washed with ethanol.
The filtrate and washings were concentrated in vacuo and the residue was dissolved in
g dichloromethane (30 mL) and treated with DIEA (500 mL). The solution was added dropwise to
cis-1,5-cyclooctylene di(chloroforrnate) (4.15g; 15.4 mmol), prepared as in Example 5, in ice
cold dichloromethane (75 mL). The reaction mixture was stirred overnight and diluted with 0.5N
12 aqueous KHSO4 and dichloromethane. The aqueous phase was extracted with dichloromethane
and the combined organic layers were washed with brine, dried (Na2SO4) and concentrated.
Purifying from the residue by chromatography on silica gel (ethyl acetate-hexane, 3: 1; then pure
15 ethyl acetate) gave cis-1,5-cyclooctylene chloroformate
4-[2-(4-tert-butoxycarbonylpiperidin-4-yl)ethylcarbarnoyl]-l-piperazinecarboxylate (1.02 g;
1.8 rnmol, 42%) as a pale yellow oil; 'H-NMR (300 MHz, CDC13): 5.00-4.90 (m, lH), 4.85-4.75
18 (m, lH), 4.35 (br t, lH), 4.05 (br d, 2H), 3.50-3.40 (m, 4H), 3.35-3.30 (m, 4H), 3.25 (q, 2H), 2.65
(t, 2H), 2.05-1.55 (m, 17H), 1.40 (s, 9H), 1.25-1.00 (m, 2H).
EXAMPLE 27
21 4-Cyanophenylacetic acid
2-(4-Cyanophenyl)ethanol (5.00 g; 34.0 mmol) was dissolved in acetone (140 mL) and
cooled to 10-1~~C. A solution of CrO3 in aqueous H2SO4 was added dropwise, while keeping the
24 internal ~e~ ,e~dlure below 30~C, until an orange color persisted giving a suspension. The
suspension was stirred for 45 mmutes and filtered. The solids were washed with acetone
(150 mL) and the combined filtrate and washings were stirred with 2-propanol (20 mL ) for 30
27 minutes. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was
,
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taken intoethyl acetate and the solution was washed with 0.5N aqueous KHSO4, water and brine
and concentrated. The residue was dissolved in dichloromethane and the solution was treated
3 with sodium hydroxide (1.56 g) in water (100 mL). The aqueous phase was extracted with
dichloromethane and acidified to pH 1-2 with concentrated aqueous hydrochloric acid giving a
ci~ le. The ~leci~ te was washed with water and air-dried. Drying in vacuo gave
6 4-cyanophenylacetic acid (3.36 g, 20.7 mmol, 61%) as a white powder; 'H-NMR (300 MHz,
CDCI3): 7.63 (d, 2H), 7.40 (d, 2H), 3.72 (s, 2H).
EXAMPLE 28
9 ter~-Butyl 4-(4-cyanophenylacetyl)-1-pipela~ ecarboxylate
A mixture of 4-cyanophenylacetic acid (890 mg; 5.52 mrnol), prepared as in Example 27,
ethylene dichloride (1.16 g; 6.07 mmol) and 1-hydroxybenzotria~ole hydrate (820 mg, 6.07
12 mmol) was suspended in THF (18 mL) and tert-butyl I-piper~7.inec~rboxylate (1.04 g; 5.60
mmol) and DIEA were added to give a homogenous solution. The solution was concentrated in
vacuo and the residue was treated with 0.2N KHSO4. The mixture was filtered and the solid
5 collected was washed with water. Drying in vacuo gave tert-butyl 4-(4-cyanophenylacetyl)-
I-piperazinecarboxylate (1.68 g; 5.1 mrnol, 92%) as a solid; ~H-NMR (300 MHz, CDCl3): 7.70
(d, 2H), 7.35 (d, 2H), 3.80 (s, 2H), 3.70-3.60 (m, 2H), 3.45-3.30 (m, 6H), 1.40 (s, 9H).
18 EXAMPLE 29
tert-Butyl 4-[4-(N-hydroxyarnidino)phenylacetyl]-l-piper~7.in~cs~rboxylate
tert-Butyl 4-(4-cyanophenylacetyl)-1-pi~ dzillecarboxylate (1.68 g; 5.1 mrnol), prepared
21 as in Example 28, in dry ethanol (10 mL) was treated with hydroxylamine hydrochloride (461
mg; 6.63 mmol) and triethylamine (924 mL, 671 mg; 6.63 mmol). The mixture was heated at
reflux for 3.5 hours, cooled to room temperature and concentrated in vacuo. The residue was
24 dissolved in ethanol, filtered and the filtrate cooled overnight giving a crystalline product. The
crystals were isolated by filteration and washed with cold ethanol. Air-drying gave tert-butyl
4-[4-(N-hydroxyamidino)phenylacetyl]-l-piper~7inec~rboxylate (1.62 g; 4.5 mmol, 88%);
_, . ~
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'H-NMR (300 MHz, DMSO-d6): 9.60 (s, IH) 7.60 (d, 2H), 7.20 (d, 2H), 5.75 (s, 2H), 3.70 (s,
2H), 3.40 (br s, 4H), 3.25 (br s, 4H), 1.40 (s, 9H).
3 EXAMPLE 30
4-~Jipe~ in- l -ylcarbonylmethylbenzamidine bis(trifluoroacetate)
ter~-Butyl 4-[4-(N-hydroxyamidino)phenylacetyl]-1-piperazinecarboxylate (653 mg;6 1.81 mmol), prepared as in Example 29, and 10% palladium-on-carbon (200 mg) were suspended
in acetic acid (12 mL) and hydrogen was bubbled through the suspension overnight. The
reaction mixture was filtered and the catalyst was washed with acetic acid. The combined filtrate
g and washings were concentrated in vac~lo and the residue was dissolved in TFA. The solution
stood for 1 hour and then was concentrated in vacuo. The residue was co-evaporated from a
lllixlu~e of dichloromethane and methanol and then suspended in diethyl ether. The particulate
12 matter was collected by filtration. Drying gave 4-piperazin- 1 -ylcarbonylmethylbenzarnidine
bis(trifluoroacetate) (1.04 g; 1.81 mmol, 100%) as awhite solid; 'H-NMR (300 MHz,
DMSO-d6): 9.30 (d, 4H), 9.15 (br s, 2H), 7.70 (d, 2H), 7.40 (d, 2H), 3.85 (s, 2H), 3.65 (br d, 4H),
4.20-3.90 (m, 4H).
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EXAMPLE 3 1
cis- 1 ,5-Cyclooctylene 4-(4-amidinophenylacetyl)- 1 -pip~ hlecarboxylate
3 4-(2-piperid-4-ylethylcarbamoyl)- 1 -piperazinecarboxylate
- (Compound 94)
The following is the l)repalation of a compound of Formula I in which R' is
6 4-amidinobenzyl, R2 is 2-piperid-4-ylethyl, X' is -C(O)- and X9 is -NHC(O)-, x2 and x8 each are
1,4-piperazinylene, X3 and X7 each are -C(O)O-, X4 and x6 each are a covalent bond and X5 iS
cis- 1 ,5-cyclooctylene.
9 4-Piperazin-l-ylcarbonylmethylbenzamidine bis(trifluoroacetate) (80 mg; 0.17 rnmol),
~--,pa ed as in Example 30, was dissolved in DMF (1.0 mL) and the solution was treated with
DIEA (150 mL). cis-1,5-Cyclooctylene chloroforrnate
12 4-[2-(4-tert-butoxycarbonylpiperazin-1-yl)ethylcarbamoyl]-1-piperazinecarboxylate (100 mg),
prepared as in Example 26, in DMF (1.0 mL) was added and the mixture was stirred overnight
and concentrated in vacuo. The residue was dissolved in dichloromethane and TFA (1 :1) and the
mixture was concentrated in vacuo. The residue was triturated with diethyl ether giYing a foam
residue. Purifying from the residue by plepaldlive reverse phase HPLC and Iyophilization of the
pure fractions gave cis-1,5-cyclooctylene 4-(4-amidinophenylacetyl)-1-piperazinecarboxylate
8 4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate as a colorless solid; Electrospray
LRMS: Calculated for C35H54N8O8: MH+: 683.9; Mll2t2/2: 342.5" Found: MH+: 683.8;MH2+2/2: 342.3.
21 Proceeding as in Example 31 and substituting different starting materials the following
compounds of Formula I were prepared:
cis-1,5-cyclooctylene 4-(1-amidinopiperid-4-ylacetyl)-1-piperazinecarboxylate
24 4-(2-piperid-4-ylethylcarbarnoyl)-1-piperazinecarboxylate (Compound 95); Calculated for
C34H59N9O6: MH~: 690.9, Found: MH+: 690.6;
cis- 1,5-cyclooctylene 4-(4-~m~ nobenzoylaminomethyl)-1-piperidinecarboxylate
27 4-(2-piperid-4-ylethylcarbarnoyl)-1-piperazinecarboxylate (Compound 96); Calculated for
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C36Hs6N8O6: MH+: 697.9, Found: MH+: 697.7;
cis-l,S-cyclooctylene 4-(4-amidinobenzylcarbamoyl)-l-piper~7.inec~rboxylate
3 4-(2-piperid-4-yl)ethylcarbamoyl]~ ip~ iLIecarboxylate (Compound 97); Calculated for
C35H55NgO6 MHt: 698.9, Found: MH+: 698.7;
cis-1,5-cyclooctylene 4-(4-amidinophenylsulfonylaminomethyl)-1-piperidinecarboxylate
6 4-(2-piperid-4-ylethylcarbamoyl)-1-piper~7inec~rboxylate (Compound 98); Calculated for
C35H56N8O7: MH+: 733.9, Found: MH+: 733.4;
cis- 1,5 -cyclooctylene 4- ~2-(1 -amidinopiperid-4-yl)ethylcarbamoyl] -
9 I-piperazinecarboxylate 4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate
(Compound 99); Calculated ~or C35H62N~0O6: MH+: 719.9, Found: MH+: 719.5;
cis- l ,S-cyclooctylene 4-(4-amidinophenylcarbamoylmethyl )- I -piperidinecarboxylate
2 4-(2-piperid-4-ylethylcarbamoyl)- 1 -piperazinecarboxylate (Compound 100); Calculated for
C36H56N8O6: MH+: 697.9, Found: MH+: 695.6;
cis-l,S-cyclooctylene 4-(4-N-methoxycarbonylamidinobenzylcarbamoyl)-
15 I-piperazinecarboxylate 4-(2-piperid-4-ylethylcarbamoyl)-1-piperazinecarboxylate
(Compound 101); Calculated for C37H57NgO8 MH+: 756.9, Found: MH+: 756.4; and
cis-1,5-cyclooctylene 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate
8 3-piperid-4-ylpropyl-1-piperidinecarboxylate (Compound 102); Calculated for
C36Hs8N8O5: MH+: 683.9 Found: MH+: 683.3.
EXAMPLE 32
21 l ,S-Pentamethylene di [4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate]
(Compound 103)
The following is the ~ )al~lion of a compound of Formula I in which R' and R2 each are
24 4-guanidinobenzyl, X' and Xg each are -NHC(O)-, x2 and X~ each are 1,4-piperazinylene, X3 and
X' each are -C(O)O- and X4-X6-X5 together are l,S-pentamethylene.
~ert-Butyl 4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate trifluoroacetate
27 (306 mg, 0.62 mmol.) was treated with neat trifluoroacetic acid (lmL) at room temperature over
ten mimlte~ to give a colorless homogeneous solution. The liquid was concentrated and the oily
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residue triturated with diethyl ether (3x lOmL) followed by drying in l~acuo to a colorless foam.
The deprotected piperazine salt was then taken into DMF (2.5 mL) followed by addition of
3 diisopropylethylamine (0.5 mL, 3.1 mmol.) and 1,5-n-pentylene di(chloroformate) (70 mg,
0.31 rnmol.) and the mixture was allowed to stir for one hour at room temperature. The reaction
mixture was concentrated and the residue was triturated with diethyl ether (3x 1 OmL) followed
6 by drying in vucuo. The crude material was taken into water (5 mL) and purified by prepa~a~ive
reverse phase HPLC and Iyophilization to give 1,5-n-pentylene
di~4-(4-guanidinobenzylcarbamoyl)-1-piperazinecarboxylate] as a colorless amorphous solid;
g Electrospray LRMS: Calculated for C33H48N,206: MH+: 709.8" Found: MH+: 709.3.
Proceeding as in Example 32 and substituting different starting materials the following
compounds of formula I were prepared:
1 2 1,4-tetramethylene di [4-(4-guanidinobenzylcarbamoyl)- 1 -piperazinecarboxylate]
(Compound 104); Calculated for C3~H46N,206: MH+: 695.8, Found: MH+: 695.8;
4-guanidinobenzyl
15 4- { 5- [4-(4-guanidinobenzylcarbarnoyl)piperazin- 1 -ylcarbonyl]valeryl } - 1 -piperazinecarboxamide
(Compound 105); Calculated for C32H46N,204: MH+: 663.8, Found: MH+: 663.4;
4-guanidinobenzyl
18 4-{6-[4-(4-guanidinobenzylcarbamoyl)piperazin-1-ylcarbonyl]hexanoyl}-
l-pilJt;ld~i~lecarboxamide (Compound 106); Calculated for C33H48N,204: MH+: 677.8,
Found: MH+: 677.4;
21 4-guanidinobenzyl
4- { 7-[4-(4-guanidinobenzylcarbarnoyl)piperazin- 1 -ylcarbonyl]heptanoyl } -
1-piperazinecarboxamide (Compound 107); Calculated for C34H50NI204: MH+: 691.9,
24 Found: MH+: 691.5;
4-guanidinobenzyl
~ 4- { 8-[4-(4-guanidinobenzylcarbamoyl)piper~ill- 1 -ylcarbonyl]octanoyl } -
27 l-pip~l~Ginecarboxarnide (Compound 108); Calculated for C37H57Ng08 MHt: 756.9,
Found: MHt: 756.4;
,
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4-guanidinobenzyl
4- { 9-[4-(4-guanidinobenzylcarbamoyl)piperazin- 1 -ylcarbonyl lnonanoyl } -
3 1-piperazinecarboxamide (Compound 109); Calculated for C36H54N,2O4: MH+: 719.9,
Found: MH+: 719.5;
4-amidinobenzyl
6 4-{7-[4-(4-amidinobenzylcarbamoyl)piperazin-1-ylcarbonyl]heptanoyl}-
1-piperazinecarboxamide (Compound 110); Calculated for C34H48N,0O4: MH+: 661.8,
Found: MH+: 661.3;
9 1,5-pentamethylene di[4-(4-guanidinophenylacetyl)piperazin-1-ylcarbonyl](Compound 111); Calculated for C33H46N,0O4: MH+: 647.8, Found: MH+: 647.3;
1,6-hexamethylene di[4-(4-guanidinophenylacetyl)piperazin-1-ylcarbonyl]
12 (Compound 112); Calculated for C34H48N,0O4: MH~: 661.8, Found: MH+: 661;
1,7-heptamethylene di [4-(4-guanidinophenylacetyl)piperazin- 1 -ylcarbonyl]
(Compoundll3);CalculatedforC35HsON,0O4: MH+: 675.9,Found: MH~: 675.4;and
3-oxa- 1,5-pentamethylene di[4-(4-guanidinophenylacetyl)piperazin- 1 -ylcarbonyl]
(Compound 114); Calculated for C32H44N,0O7: MH+: 681.8, Found: MH+: 681.4.
E~AMPLE 32
18 In Vitro Assay of Tryptase Inhibition
Mixtures of human tryptase (15 ~lg/mL) and test compound (varying concentrations) in
Tris buffer (comprising: NaCl, 100 mM; Tris, 50 mM; 2-[N-morpholine]ethane sulfonic acid,
21 2.5 mM, CaCI2, 0.5 mM; DMSO, 10%; glycerol, 5%; polyoxyethylenesorbitan monolaurate
(Tween-20), 0.05%; heparin, 25 ng/mL; and pH 8.2) were incubated for I hour at room
temperature and then Tosyl-Gly-Pro-Lys-para-nitroanilide was added such that the final
24 concentration of the assay mixture was 0.5 mM. Hydrolysis of the substrate was followed
spectrophotometrically at (405 nm) for 5 minutes. Apparent inhibition constants (Ki) were
calculated from the enzyme progress curves using standard mathematical models.
27 Human tryptase can be purified from human lung and skin tissue samples (e.g., see
Smith etal. (1984) J.Biol. Chem. 59: 11046-11051,andBraganza etal.(1991)Biochem.
. ..
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30:4997-5007) and human mast cell line or obtained commerically (e.g., ICN Biomedicals,
I~ine California; Athens Research & Technology, Athens, Georgia). Porcine intestinal mucosa
3 heparin and Tosyl-Gly-Pro-Lys-para-nitroanilide can be obtained from Sigma Chemical
Company.
Proceeding as described in this application or by methods known to those of ordinary
6 skill the following compounds of Formula I were prepared and tested for tryptase inhibitory
activity:
Compound l, Kj=0.003~ M; Compound 2, Kj=0.8~ M; Compound 3, Kj=0.07~ M;
9 Compound 4, Kj=0.001~ M; Compound 5, Kj=0.2~ M; Compound 6, Kj=l~ M;
Compound 7, Kj=0.3~ M; Compound 8, Kj=4~ M; Compound 9, Kj=0.4~ M;
Compound l 0, Kj=l~ M; Compound 1 l, Kj=0.09~ M; Compound 12, Kj=0.2~ M;
12 Compound 13, Kj=0.02~ M; Compound 14, Kj=0.004~ M; Compound lS, Kj=0.5~ M;
Compound 16, Kj=0.9~ M; Compound 17, Kj=l~ M; Compound 18, Kj=0.08~ M;
Compound l9, Kj=1.2~ M; Compound 20, Kj=3.4~ M; Compound 21, Kj=0.5~ M;
5 Compound 22, Kj=0.2~ M; Compound 23, Kj=4~ M; Compound 24, Kj=0.3~ M;
Compound 25, K;=0.002~ M; Compound 26, Kj=19~ M; Compound 27, Kj=2~ M;
Compound 28, Kj=4~ M; Compound 29, Kj=l~ M; Compound 30, Kj=0.031~ M;
18 Compound 31, Kj=l~ M; Compound 32, Kj=2~ M; Compound 33, Kj=l~ M;
Compound 34, Kj=3~ M; Compound 35, Kj=0.8~ M; Compound 36, Kj=0.6~ M;
Compound 37, Kj=0.07~ M; Compound 38, Kj=0.004~ M; Compound 39, Kj=0.004~ M;
21 Compound 40, Kj=4~ M; Compound 41, Kj=0.7~ M; Compound 42, Kj=0.02~ M;
Compound 43, Kj=0.4~ M; Compound 44, Kj=0.02~ M; Compound 45~ Kj=0.08~ M;
Compound 46, Kj=l~ M; Compound 47, Kj=0.3~ M; Compound 48, Kj=0.09~ M;
24 Compound 49, Kj=2~ M; Compound 50, Kj=0.08~ M; Compound 51, Kj=l~ M;
Compound 52, Kj=0.04~ M; Compound 53, Kj=6~ M; Compound 54, Kj=0.1~ M;
Compound 55, Kj=2~ M; Compound 56, Kj=10~ M; Compound 57, Kj=2~ M;
27 Compound 58, Kj=0.1~ M; Compound 59, Kj=0.5~ M; Compound 60, Kj=5~ M;
Compound 61, Kj=41~ M; Compound 62, Kj=0.2~ M; Compound 63, Kj=2~ M;
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Compound 64, Kj=1 ~M; Compound 65, Kj=0.001 ~lM; Compound 66, Kj=0.02~LM;
Compound 67, Kj=3,uM; Compound 68, Kj=0.04~M; Compound 69, Kj=0.511M;
3 Compound 70, Kj=0.0511M; Compound 71, Kj-0.811M; Compound 72, Kj=O.l~lM;
Compound 73, Kj=0.002~M; Compound 74, Kj=0.0411M; Compound 75, Kj=0.01~1M;
Compound 76, Kj=0.1!1M; Compound 77, Kj=6~M; Compound 78, Kj=0.1,uM;
6 Compound 79, K,=IIlM; Compound 84, Kj=0.06~LM; Compound 85, Kj=O.9,uM;
Compound 86, Kj=0.08~M; Compound 87, Kj=0.05,uM; Compound 88, Kj=0.1 ~IM;
Compound 89, Kj=O.l,uM; Compound 90, Kj=1!1M; Compound 91, Kj=0.1~1M;
9 Compound 92, Kj=0.1~M; Compound 93, Kj=0.02~M; Compound 94, Kj=0.007~LM;
Compound 95, Kj=0.0211M; Compound 96, Kj=0.02~M; Compound 97, Kj=0.0009~1M;
Compound 98, Kj=0.0311M; Compound 99, Kj=0.05~LM; Compound 100, Kj=0.00911M;
2 Compound 101, Kj=0.04~M; Compound 102, Kj=0.08~M; Compound 103, Kj=0.00111M;
Compound 104, Kj=0.003,uM; Compound 105, Kj=0.04~M; Compound 106, Kj=0.004~1M;
Compound 107, Kj=0.0001,uM; Compound 108, Kj=0.000511M; Compound 109 Kj=0.0007~M;
15 Compound 110, Kj=0.000811M; Compound 111, Kj=0.3~M; Compound 112, Kj=O.O9~M;
Compound 113 Kj=O.OO5~lM; and Compound 114, Kj=0.05~M.
EXAMPLE 33
18 In Yivo Assay of Asthma
Allergic sheep characterized as dual responders (i.e., displaying early and late phases of
bronchoconstriction) are challenged with antigen (e.g., Ascaris suum). The sheep are
21 ~-lmini~tered test compound or vehicle by aerosol inhalation at 0.5 hours before and at 4 and 24
hours post antigen challenge. Specific lung resistance (SRL) is monitored via an esophageal
balloon catheter just prior to the first test compound or vehicle treatment and every 0.5 to 1 hour
24 thereafter.
In addition, airway responsiveness is monitored 1 to 2 days prior to antigen challenge and
just subsequent to arln~ini~tration of test compound or vehicle at 24 hours post antigen challenge.
27 For the purposes of this application, airway responsiveness is defined as the cumulative dose of
carbachol required to increase SRL by 400% (PC400). The PC400 values are obtained by
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a-lmini.ctering 0 to 30 breath units of 1% carbachol (l 0 mg in I mL of PBS) by aerosol inhalation
until SRL was increased by 400%.
3 Sheep treated with vehicle exhibit early phase bronchoconstriction from 0 to 4 hours post
antigen challenge and late phase bronchoconstriction from 4 to greater than 8 hours post antigen
challenge. In addition, vehicle treated sheep exhibit hyper responsiveness to carbachol (i.e., a
6 60% decrease in PC400 is observed).
Sheep treated with tryptase inhibitors do not exhibit late phase broncoconstriction (i.e., at
4 to 8 hours post antigen challenge, SRL remained at basal levels). Further, sheep treated with
g tryptase inhibitors do not exhibit any hyper responsiveness to carbachol.
EXAMPLE 34
Representative Pharmaceutical Formulations Containing a Compound of Formula 1.
12 ORAL FORMULATION
Compound of Formula I l 0- l 00 mg
Citric Acid Monohydrate l 05 mg
Sodium Hydroxide l 8 mg
Flavoring
Water q.s. to l 00 mL
18 INTRAVENOUS FORMULATION
Compound of Formula I 0.1 -l 0 mg
Dextrose Monohydrate q.s. to make isotonic
21 Citric Acid Monohydrate l.05 mg
Sodium Hydroxide 0. l 8 mg
Water for Injection q.s. to l.0 mL
24
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TABLET FORMULATION
Compound of Formula I 1%
3 Microcrystalline Cellulose 73%
Stearic Acid 25%
Colloidal Silica 1%.
